Abstract：Alzheimer's disease （AD） is a deleterious neurodegenerative disorder， which has become a significant public health concern and economic burden. The pathogenesis of AD is complex and involves several hypotheses such as amyloid β-protein （Aβ） deposition， Tau protein hyperphosphorylation， oxidative stress， and inflammation. There is an urgent need for a holism-based comprehensive intervention with multi-pathway， multi-level， and multi-target characteristics， which demonstrates the unique advantage of traditional Chinese medicine （TCM）. Therefore， it is of great significance to conduct and promote research on TCM treatment of AD. Danggui Shaoyaosan （DSS） from the Synopsis of Golden Chamber （《金匮要略》） by ZHANG Zhongjing （150 AD-219 AD） was originally designed for reliving gynecological ailments. It is a classic TCM formula that modulates liver and spleen and dispels blood stasis and water retention. Since the late 1980s when Japanese researchers reported its therapeutic effect on AD， it has been widely used in the clinic with clear effects. The elucidation of the mechanism of this formula helps exert its effects. Hereby， this paper reviewed relative research progress and made an analysis in terms of attenuating aberrant accumulation of Aβ and hyperphosphorylated Tau protein， anti-inflammatory and antioxidant activities， mediating neurotransmitters， ameliorating lipid metabolism， modulating gut microbiota， reduced neuron apoptosis， decreasing intracellular Ca2+ overloading， and increasing the expression of estradiol. This paper is expected to provide references for understanding the scientific connotation of DDS in the treatment of AD and lay a solid foundation for further investigation.
Abstract：ObjectiveTo investigate the mechanism of Danggui Shaoyaosan （DSS） in the improvement of the cognitive ability of SAMP8 mice with Alzheimer's disease （AD） via regulating the ubiquitin-proteasome pathway （UPP）.MethodFifteen SAMR1 mice were used as a normal group， and 60 SAMP8 mice were randomly divided into a model group and DSS high， medium， and low-dose groups （57.6， 28.8， and 14.4 g·kg-1·d-1）， with 15 mice in each group. Intragastric administration was conducted for eight continuous weeks. Place navigation and spatial capacity were evaluated by Morris water maze. Pathological structure changes in neurons in the hippocampal CA1 area was detected by hematoxylin-eosin （HE） staining. The protein expression levels of hippocampal β-amyloid protein（Aβ） and phosphorylation（p）-Tau were determined by immunohistochemical staining （IHC） and enzyme-linked immunosorbent assay （ELISA）， and the mRNA and protein expression levels of hippocampal ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase-1 （UCHL1）， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.ResultAs compared with the normal group， the escape latency was prolonged in the model group （P<0.05） with the reduced number of crossing platform quadrants and time ratio in the platform quadrant （P<0.05）. The model group decreased neurons and condensed cell bodies in the CA1 area， and increased β-amyloid precursor protein （β-APP） and p-Tau positive cells （P<0.05）. In the model group， the protein expression levels of Aβ and p-Tau were increased （P<0.05）， the mRNA and protein expression levels of Ub were increased （P<0.05）， and the mRNA and protein expression levels of E3， 26S proteasome， UCHL1， and UCHL3 were decreased （P<0.05）. As compared with the model group， the escape latency was shortened in the DSS high and medium-dose groups （P<0.05） with an increased number of crossing platform quadrants and residence time ratio （P<0.05）. The pathological changes in CA1 of each DSS group were significantly improved， and the number of β-APP positive staining cells decreased （P<0.05）. The number of p-Tau positive staining cells decreased in the DDS medium and low-dose groups （P<0.05）. The protein expression levels of Aβ and p-Tau in each DDS group decreased （P<0.05）， and the mRNA expression level of Ub in each group decreased （P <0.05）. The mRNA expression levels of 26S， E3， and UCHL3 in the DDS high and medium-dose groups increased （P<0.05）， and the mRNA expression level of UCHL1 in the DDS medium-dose group increased （P<0.05）. The protein expression level of Ub in each DDS group decreased， and the protein expression levels of 26S， E3， UCHL1+3 in the DDS high and medium-dose groups increased （P<0.05）.ConclusionDSS can improve the cognitive ability of SAMP8 mice， and its mechanism may be related to the reduction of the abnormal deposition of Aβ and p-Tau via decreasing the expression of Ub and increasing that of E3， 26S， UCHL1， and UCHL3 in the UPP.
Abstract：ObjectiveTo investigate the protective effect of Danggui Shaoyaosan （DSS）-contained serum on β-amyloid （Aβ）1-40-injured rat adrenal pheochromocytoma PC12 cells and its mechanism in regulating ubiquitin-proteasome pathway （UPP）.MethodAβ1-40 was used to intervene PC12 cells to prepare the cell models of Alzheimer's disease （AD）， and the experiment was divided into the blank， model， and DSS-contained serum high， medium， and low-dose groups （10%， 5%， and 2.5%）. Cell viability and apoptosis were detected using cell counting kit-8 （CCK-8） method and flow cytometry， respectively. The content of Aβ and p-Tau protein was determined by enzyme-linked immunosorbent assay （ELISA）. The ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase1 （UCHL1）， and UCHL3 protein expressions of UPP were displayed using immunofluorescence cytochemistry （ICC）， and the mRNA and protein expression levels of Ub， E3-parkin， 26S， UCHL1， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.ResultThe data of the CCK8 experiment verified that 5 μmol·L-1 and 48 hours were the optimal conditions for modeling Aβ1-40-injured PC12 cells. As compared with the blank group， the cell viability rate in the model group decreased （P<0.05） with an increased apoptosis rate （P<0.05）， the content of Aβ and p-Tau contents was elevated （P<0.05）， the mRNA and protein expression levels of Ub increased， and the mRNA and protein expression levels of 26S， E3， and UCHL1+3 decreased （P<0.05）. As compared with the model group， the cell viability rate in the DSS-contained medium-dose group increased （P<0.05）， whereas the apoptosis rate in each DSS-contained group decreased （P<0.05）. The content of Aβ in each DDS-contained group decreased （P<0.05）， and the content of p-Tau in the DDS-contained high and medium-dose groups decreased （P<0.05）. The mRNA expression level of Ub decreased， and that of 26S increased in each DDS-contained group （P<0.05）. The mRNA expression level of UCHL1 in the DDS-contained medium-dose group increased （P<0.05）， and the mRNA expression levels of E3 and UCHL 3 in the DDS-contained high and medium-dose groups increased （P<0.05）. The protein expression level of Ub in each DDS-contained group decreased， and the protein expression levels of 26S， E3， and UCHL1+3 in the DDS high and medium-dose groups increased. The DSS-contained serum medium-dose group exerted the optimal effect.ConclusionDSS-contained serum can increase cell viability rate， reduce cell apoptosis rate， eliminate Aβ and p-Tau protein deposits， and exert protective effects on Aβ1-40-injured PC12 cells. Its mechanism may involve UPP via decreasing the expression of Ub and increasing that of 26S， E3， UCHL1， and UCHL3.
Abstract：ObjectiveTo analyze the effects of Danggui Shaoyaosan （DSS） on the gut microbiota of the Alzheimer's disease （AD） model in SAMP8 mice based on 16S rDNA sequencing.MethodTwenty-four SAMP8 mice aged seven months were randomly divided into low-， medium-， and high-dose DSS groups （14.4， 28.8， 57.6 g·kg-1·d-1） and a model group according to a random number table， with six rats in each group. Six SAMR1 mice of the same age were assigned to the normal group. After intragastric administration for eight consecutive weeks， 16S rDNA sequencing was performed to detect the gut microbiota of feces in mice. Morris water maze was employed to assess the directional navigation and space exploration ability of mice. Nissl staining was performed to observe the pathological changes of neurons in the hippocampal CA1 area. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the protein content of hippocampal amyloid β-protein （Aβ） and hyperphosphorylated Tau （p-Tau）.ResultCompared with the normal group， the model group presented a declining α diversity （P<0.05）， markedly altered β diversity， prolonged escape latency （P<0.05）， reduced number of platform crossings and cumulative duration in the targeted quadrant （P<0.05）， decreased neurons and Nissl bodies in the CA1 hippocampal area， and up-regulated Aβ and p-Tau expression （P<0.05）. However， DSS intervention enhanced the α diversity， and medium- and high-dose DSS， especially the medium-dose DSS， could result in α diversity similar to the control group. Moreover， at the phylum level， the abundance of Firmicutes increased （P<0.05）， while the abundance of Bacteroidetes and Proteobacteria decreased （P<0.05）. At the genus level， the abundance of Lactobacillus and other genera increased （P<0.05）， while the abundance of Bacteroides， Helicobacterium， Rikenella， Parabacteroides， Sutterella， and Mucilaginibacter decreased （P<0.05）. The DSS groups also showed shortened escape latency （P<0.05）， increased number of platform crossings and cumulative duration in the targeted quadrant （P<0.05）， increased Nissl bodies （P<0.05）， and reduced Aβ and p-Tau content （P<0.05）. Pearson correlation analysis showed that the abundance of Mucilaginibacter， Bacteroides， and Sutterella was negatively correlated with the cognitive ability of SAMP8 mice， while the abundance of Lactobacillus and Butyricimonas was positively correlated with the cognitive ability of SAMP8 mice.ConclusionDSS can improve the cognitive ability of SAMP8 mice， and its mechanism may be related to the regulation of gut microbiota diversity and community composition.
Abstract：ObjectiveTo study the expression changes of Lon protein and mitochondrial dynamics-related protein in the hippocampus of SAMP8 mice and provide a theoretical basis for the treatment of Alzheimer's disease by invigorating the spleen and supplementing Qi.MethodEight 3-month-old SAMR1 mice were used as the normal group， and 32 3-month-old SAPM8 mice were divided into model group， western medicine group （0.013 g·kg-1）， low-dose Si Junziwan group （3.24 g·kg-1）， and high-dose Si Junziwan group （12.56 g·kg-1）， with 8 mice in each group. The western medicine group was gavaged with donepezil， and the Si Junziwan low- and high-dose groups were gavaged with Si Junziwan for 30 days. The positioning navigation experiment of the water maze was started on the 25th day， and the space exploration experiment of the water maze was started on the 30th day. On the 30th day， the protein expression of mitofusin 2 （MFN2） was detected by immunohistochemistry， the expression of adenosine 5'-monophosphate （AMP）-activated protein kinase （AMPK） was detected by enzyme-linked immunosorbent assay （ELISA）， the content of ATP was detected by colorimetry， the microstructure of neuron mitochondria was detected by electron microscope， and the expressions of Aβ protein， Lon protein， dynamin-related protein 1 （DRP1） protein， and MFN1 protein were detected by Western blot.ResultAs compared with the normal group， the latency escape period increased， the number of crossings decreased， the expression of AMPK increased， and the content of ATP decreased in the model group. The expressions of Aβ protein and DRP1 protein increased （P<0.01）， whereas the expressions of Lon protein， MFN1 protein decreased in the model group （P<0.05，P<0.01）， and MFN2 protein decreased. The vacuolation of mitochondria increased and the cristae broke in the model group. As compared with model group， the time of the latent escape period decreased （P<0.01）， and the number of crossings increased in the low-dose and high-dose Si Junziwan groups （P<0.05）. The expression of AMPK （P<0.01） decreased， the content of ATP increased （P<0.01）， the expression of Aβ and DRP1 protein decreased （P<0.05， P<0.01）， and the expression of MFN1 protein was up-regulated （P<0.05） in high-dose Si Junziwan groups. The vacuolation was more obvious in the low-dose Si Junziwan group， whereas the vacuolation was restored and the ridge was clear in the high-dose Si Junziwan group.ConclusionSi Junziwan treats Alzheimer's disease by up-regulating the protein expression of Lon， correcting the disorder of mitochondrial division and fusion protein， and changing the memory function of SAMP8 mice.
Abstract：ObjectiveTo evaluate the pharmacological effect of Alismatis Rhizoma （AR） and its processed product on rats with edema of kidney Yin deficiency and explore the mechanism.MethodA total of 42 male SPF SD rats were randomized into normal group （equivalent volume of distilled water）， model group （equivalent volume of distilled water）， positive medicine Liuwei Diguangwan group （1.4 g·kg-1）， low- and high-dose AR groups （1， 4 g·kg-1， respectively）， and low- and high-dose salt-processed AR （SAR） groups （1， 4 g·kg-1， respectively）， with six rats in each group. Adriamycin （tail vein injection） and thyroxine （gavage） were used to induce edema of kidney Yin deficiency in rats except the normal group. The administration lasted 4 weeks for all the groups. After the last administration， histopathological changes of rat kidneys were observed based on hematoxylin-eosin （HE） staining. Serum content of triiodothyronine （T3）， thyroxine （T4）， follicle-stimulating hormone （FSH）， and testosterone （T） was determined by radioimmunoassay， and serum content of creatinine （CREA）， urea （UREA），cholesterol （CHOL） and triglyceride （TG） by automatic biochemical analyser. The levels of gonadotropin-releasing hormone （GnRH）， cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） in plasma were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of aquaporin（AQP）-1 and AQP-2 and the transcription of mRNA in kidney were measured by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， respectively.ResultCompared with normal group， the rats in model group showed decrease in body mass and urine volume （P<0.01）， increase in water consumption （P<0.05）， infiltration of a large number of inflammatory cells and fibrous tissue proliferation in the kidney， rise of the expression and transcript levels of T3， T4， cAMP/cGMP， CREA， FSH， AQP-1， and AQP-2 （P<0.01）， the contents of CHOL and TG were significantly increased （P<0.05）， and reduction in the levels of GnRH and T （P<0.01）. Body mass increased in both the low- and high- dose groups of AR and SAR compared with that in model group， with significant differences between the low-dose AR group and the low-dose SAR group （P<0.01）. Moreover， compared with model group， low- and high-dose AR and SAR insignificantly increased the urine volume of rats， reduced the inflammatory cells in kidney tissues， significantly decreased the levels of T4， cAMP/cGMP， UREA， CREA， FSH， CHOL and TG in serum （P<0.05，P<0.01）， and elevated the level of GnRH （P<0.01）， high-dose AR， low- and high-dose SAR significantly lowered the transcription levels of AQP-1 and AQP-2 mRNA in the kidneys of rats （P<0.01）.ConclusionBoth AR and SAR alleviated the edema of kidney Yin deficiency in rats by down-regulating the expression of AQP-1 and AQP-2 and correcting the hypothalamic-pituitary-gonadal （HPG） axis disorder.
Abstract：ObjectiveTo investigate the preventive effect and mechanism of Aurantii Fructus Immaturus and naringin on postoperative intestinal adhesion in rats.MethodThe preventive effect of Aurantii Fructus Immaturus and naringin on intestinal adhesion was studied by cecal scraping model of rats， the model rats were randomly divided into model group， dexamethasone sodium phosphate group and Aurantii Fructus Immaturus low， medium and high dose groups （1.58， 3.15， 6.30 g·kg-1·d-1）， tsham-operated group was treated with an incision in the abdomen. Adhesion was assessed by Nair method after 7 d of administration. Hematoxylin-eosin （HE） staining was used to observe the pathological morphology and inflammatory cell infiltration of cecum， the expression of matrix metalloproteinase-9 （MMP-9） in cecal adhesion was detected by immunohistochemistry. Meanwhile， intestinal adhesion model rats were randomly divided into model group， dexamethasone sodium phosphate group， naringin low， medium， high dose groups （50， 100， 200 mg·kg-1·d-1）， and the sham-operated group was treated with an incision in the abdomen. Adhesion was assessed after 7 d of administration， HE staining was used to observe the pathological morphology and inflammatory cell infiltration in the cecum， and the expression of MMP-9 in the cecal adhesion tissue was detected by immunohistochemistry. Expressions of transforming growth factor-β1 （TGF-β1） and α-smooth muscle actin （α-SMA） in cecum were analyzed by western blot.ResultAurantii Fructus Immaturus could inhibit the formation of postoperative intestinal adhesions in rats， reduce the inflammatory response of damaged cecum tissue， and up-regulate the expression of MMP-9 in the adherent tissue in a dose-dependent manner. All dose groups of naringin could significantly inhibit the formation of postoperative intestinal adhesions in rats， reduce inflammatory cell infiltration and fibrous proliferation in tissue， up-regulate the expression of MMP-9 in the adherent tissue in a dose-dependent manner， and inhibit the expression of TGF-β1 and α-SMA in cecal tissue.ConclusionAurantii Fructus Immaturus and naringin can reduce postoperative intestinal adhesion formation in rat model， and their effects may be related to reducing tissue fibrosis and accelerating extracellular matrix degradation.
Abstract：ObjectiveTo observe the effect of the serum containing Puerariae Lobatae Radix on tumor necrosis factor-α （TNF-α）， endoplasmic reticulum stress signaling pathway protein endoplasmic reticulum stress protein glucose-regulated protein 78 （GRP78）， activated transcription factor 4 （ATF4）， protein kinase R-like endoplasmic reticulum kinase （PERK）， and eukaryotic translation initiation factors （eIF2α） and on inflammatory injury of macrophages induced by high glucose， and explore its possible mechanism.MethodThe control serum and serum containing Puerariae Lobatae Radix were prepared by serum pharmacology. The RAW264.7 macrophages were cultured in vitro， and 62.5 mmol·L-1 glucose was used to induce macrophages to establish a model of severe endoplasmic reticulum （ER） stress （ERS） in vitro. Different volume fractions of serum containing Puerariae Lobatae Radix and ERS inhibitor （4-PBA） were used to interfere with the cells. Different glucose concentrations （22.5， 23.5， 25.0， 27.5， 32.5， 47.5，72.5， 122.5 mmol·L-1） and the effect of different volume fractions of serum containing Puerariae Lobatae Radix （0%， 2.5%， 5%， 7.5%， 10%， 15%， 20%） on the survival rate of RAW264.7 macrophages were detected by cell proliferation and cell counting kit-8 （CCK-8） assay. Based on the results of the effect of glucose concentrations on macrophage survival rate by CCK-8， Western blot （WB） was used to determine the protein expression levels of GRP78， the signature protein of ERS， by different concentrations of glucose （22.5， 32.5， 42.5， 52.5， 62.5， 72.5 mmol·L-1） at different time periods （6， 12， 24， 36， 48 h）， and the optimal concentration and time for establishing the model of severe ERS were screened out. Based on the above experimental results， the cells were divided into blank group， 62.5 mmol·L-1 glucose model group， 2 mmol·L-1 4-PBA group， and high， medium， and low-dose （15%， 10%， 5%） serum containing Puerariae Lobatae Radix groups for subsequent experiments. The expression of TNF-α in the supernatant of RAW264.7 macrophages in the model of severe ERS was determined by enzyme-linked immunosorbent assay （ELISA）. The expressions of related pathway proteins in the model of severe ERS were determined by WB.ResultThe results of CCK-8 assay showed that the survival rate of macrophages reached the highest under the stimulation of glucose concentration of 27.5 mmol·L-1 （P<0.01）， while the survival rate of macrophages increased with the concentration increasing from 22.5 mmol·L-1 to 27.5 mmol·L-1. When the glucose concentration was 25.0 mmol·L-1， there was a significant difference （P<0.01）， and when the glucose concentration was 37.5 mmol·L-1 to 122.5 mmol·L-1， there was a downward trend. The serum containing Puerariae Lobatae Radix showed significant differences in the volume of 1% to 15% （P<0.05， P<0.01）. The results of WB found that the GRP78 protein expression was the most significant at 24 h （P<0.01）， and the GRP78 protein expression was the most significant when the glucose concentration was 62.5 mmol·L-1 （P<0.01）. Therefore， 62.5 mmol·L-1 of glucose was the optimal concentration to induce the model of severe ERS. As compared with the blank group， the protein expression levels of TNF-α， GRP78， ATF4， phosphorylation（p）-PERK/PERK， p-eIF2α/eIF2α in the model group were significantly increased （P<0.05， P<0.01）， and as compared with the model group， the protein expression levels of TNF-α， GRP78， ATF4， p-PERK/PERK， p-eIF2α/eIF2α in each administration group were significantly decreased （P<0.05， P<0.01）.ConclusionThe results of in vitro experiments show that Puerariae Lobatae Radix can alleviate the inflammatory injury of macrophages induced by high glucose to a certain extent and restore cell homeostasis by inhibiting the expression of GRP78， ATF4， PERK， and eIF2α in the ERS signaling pathway.
Abstract：ObjectiveTo observe the effect of intranasal ginsenoside Rb1 against epilepsy and preliminarily explore the mechanism.MethodThe mouse model of chronic epilepsy was established by intraperitoneal injection of pentylenetetrazole （PTZ）. After successful modeling （21 d）， the epileptic mice were randomly divided into PTZ group， sodium valproate （VPA） group， and low-dose （20 mg·kg-1） and high-dose （40 mg·kg-1） ginsenoside Rb1 groups. Mice in each group were given corresponding drugs intranasally for 30 days， twice a day， and the control group was given the equal volume of normal saline. During the intranasal administration， the weight change， epilepsy latency， and epilepsy stage of the mice were recorded， and the changes in the electroencephalography （EEG） were recorded wirelessly. Neuronal Nuclei （NeuN） was used to observe the damage of neurons in cerebral cortex and hippocampus. The activation of microglia and astrocytes was observed with ionized calcium binding adapter molecule-1 （IBA-1） and glial fibrillary acidic protein （GFAP）， respectively. Glutamate （Glu） transporter-1 （GLT-1） and glutamine synthetase （GS） were used to observe the key molecular changes in Glu regulation.ResultCompared with the control group， the PTZ group decreased body weight （P<0.05，P<0.01）， shortened the epilepsy latency （P<0.01）， and increased the epilepsy stage （P<0.01）. The epileptic EEG waves were increased in the PTZ group. Compared with the PTZ group， the low and high-dose ginsenoside Rb1 groups increased body weight （P<0.05，P<0.01）， prolonged the epilepsy latency （P<0.05，P<0.01）， decreased the epilepsy stage （P<0.05，P<0.01）， and decreased epileptiform EEG waves. Immunofluorescence staining （IF） showed that ginsenoside Rb1 significantly ameliorated PTZ-induced neuronal damage （P<0.05，P<0.01） in the motor sensory area of the cerebral cortex and hippocampal CA1 area， and significantly inhibited PTZ-induced activation of microglia （P<0.05，P<0.01） and astrocytes. Further research found that ginsenoside Rb1 significantly improved the expressions of astrocytic GLT-1 （P<0.01） and GS （P<0.01） in the brains of epileptic mice.ConclusionIntranasal ginsenoside Rb1 can significantly improve the symptoms of epilepsy caused by PTZ in mice， which has a clear protective effect on neuronal damage in the brains of epileptic mice and significantly inhibits the activation of brain microglia and astrocyte activation. Its anti-epileptic mechanism may be related to the regulation of GLT-1 and GS of the key molecules of astrocyte Glu metabolism.
Abstract：ObjectiveTo investigate the material basis and mechanism of Arnebia euchroma against hepatocarcinoma by network pharmacology， and to verify the potential targets of A. euchroma against hepatocarcinoma by molecular docking and experiments.MethodThe main active ingredients of A. euchroma were collected by retrieving the literature through China National Knowledge Network （CNKI） and Traditional Chinese Medicine System Pharmacology Database and Analysis Platform（TCMSP）. The active ingredients were screened out by FAFDrug4 platform according to the pharmacokinetics （ADME） properties of the drugs. The screening compounds and liver cancer targets were collected by using several databases and analyzed by drawing Venn diagrams. The protein-protein interaction （PPI） network was constructed by Cytoscape and STRING database. DAVID database was used to perform Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of key targets. Autodock was used to perform molecular docking of targets on core pathways. Cell counting kit-8 （CCK-8） experiment was carried out to validate the activities of five naphthoquinones. Based on the predicted results in the H22 tumor-bearing mouse model， the key targets of isovalerylshikonin against hepatocarcinoma were verified by hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot assay.ResultFifty-five active ingredients and 34 targets of active ingredients against hepatocarcinoma were screened out. The active molecules with high degree values in the “drug-active ingredient-target-disease” network were mainly naphthoquinones. PPI network obtained several core targets of A. euchroma against hepatocarcinoma. Twenty-two pathways were screened out by KEGG analysis， mainly involving phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， vascular endothelial growth factor （VEGF）， tumor necrosis factor （TNF）， and other signaling pathways. The results of molecular docking showed that the five naphthoquinones had a good affinity with the targets of the PI3K/Akt signaling pathway. The results of CCK-8 and animal experiments showed that the lipid-soluble component isovalerylshikonin had good anti-cancer potential， and the high-dose group reduced the serum levels of VEGF and alpha fetoprotein （AFP） levels and elevated interferon-γ （IFN-γ） level （P<0.05， P<0.01）. The high-dose group also down-regulated phosphorylate（p）-Akt （Ser473） and B-cell lymphoma （Bcl）-2 protein expressions and up-regulated Bcl-2-antagonist of cell death （Bad） protein expression （P<0.01）.ConclusionA. euchroma can inhibit hepatocarcinoma cell proliferation and tumor angiogenesis and induce cancer cell apoptosis through the PI3K/Akt signaling pathway， which provides ideas and clues for the subsequent in-depth investigation of its specific mechanism.
Abstract：ObjectiveTo investigate the effect of Loki Zupa on airway remodeling in asthma based on ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS）combined with network pharmacology and experimental verification.MethodThe chemical constituents in Loki Zupa were identified by UPLC-MS. The potential active constituents of Loki Zupa were screened out based on literature retrieval， oral availability （OB） and drug-likeness （DL） in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） and Lipinski's rule of five in SwissADEM. The constituent targets of Loki Zupa were obtained through the SwissTargetPrediction. The relevant targets of airway remodeling in asthma were screened out from Online Mendelian Inheritance in Man（OMIM）， GeneCards， DrugBank， and DisGeNET. The STRING was used to conduct protein-protein interaction （PPI） among the main targets. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses were carried out through DAVID. Finally， an asthmatic airway remodeling model was induced by ovalbumin （OVA） in mice， followed by hematoxylin and eosin（HE）， periodic acid Schiff（PAS）， and Masson staining for the observation of the pathological conditions of lung tissues. The inflammatory cells in the bronchoalveolar lavage fluid（BALF） of mice were detected. The protein expression levels in mouse lung tissues were detected by Western blot and key signaling pathways were further determined.ResultEighty-two constituents were detected in the negative ion mode and 74 in the positive ion mode by UPLC-MS. Thirty-six candidate constituents and 578 predicted targets of Loki Zupa were screened out through network pharmacology， and 173 common targets with airway remodeling in asthma were obtained， including key compounds such as sebacic acid， pectolinarigenin， naringenin， apigenin， and potential targets such as protein kinase B1（Akt）1 and hypoxia-inducible factor 1α（HIF-1α）. As predicted by KEGG enrichment analysis， Loki Zupa mainly exerted the effect against airway remodeling in asthma through phosphatidylinositol 3-kinase （PI3K）/Akt， HIF-1α， mitogen-activated protein kinase （MAPK）， and other signaling pathways. Animal experiments showed that the compound formula of Loki Zupa could reduce the proliferation of airway goblet cells in asthmatic mice， improve the deposition of collagen under the airway epithelium， and decrease the up-regulated relative expression levels of phosphorylate（p）-Akt/Akt and HIF-1α by OVA sensitization in mice （P<0.05， P<0.01）， which was consistent with the results of network pharmacology.ConclusionUPLC-MS combined with network pharmacology was used to preliminarily clarify the chemical composition of Loki Zupa and its underlying mechanism in intervention in airway remodeling in asthma. Specifically， Loki Zupa presumably synergistically intervened in airway remodeling in asthma through key targets represented by Akt1 and HIF-1α， and multiple pathways represented by the PI3K/Akt and HIF-lα pathways， which is expected to provide ideas for further research on Loki Zupa.
Keywords：Loki Zupa;airway remodeling in asthma;network pharmacology;ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS）
Abstract：ObjectiveTo observe the effect of berberine combined with evodiamine on the migration and invasion of colorectal cancer HCT116 and RKO cells and to explore the underlying mechanism.Methodcell counting kit-8 （CCK-8） assay was used to examine the proliferation of HCT116 and RKO cells treated by berberine （30 μmol·L-1）， evodiamine （0.8 μmol·L-1）， and combination of two （30 μmol·L-1+0.8 μmol·L-1）， respectively. Scratch assay and Transwell assay were employed to detect the migration and invasion of HCT116 and RKO cells treated with berberine， evodiamine， and the combination， separately. In addition， the protein expression of epithelial cadherin （E-cadherin）， neural cadherin （N-cadherin）， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） in HCT116 and RKO cells treated with the berberine， evodiamine， and the combination was respectively measured by Western blot.ResultCompared with the blank group， berberine alone and evodiamine alone had no significant inhibitory effect on the proliferation， migration， and invasion of HCT116 and RKO cells， while the combination showed significant inhibition （P<0.01）. Berberine alone and evodiamine alone had no remarkable influence on the expression of PI3K， N-cadherin， and E-cadherin in HCT116 and RKO cells， but the combination significantly reduced the expression of PI3K and N-cadherin （P<0.01） and increased the expression of E-cadherin （P<0.01） in HCT116 and RKO cells. Evodiamine alone also significantly suppressed the expression of Akt protein in HCT116 and RKO cells （P<0.05）， but the suppression was weaker than that of the combination.ConclusionThe combination of berberine and evodiamine can significantly inhibit the migration and invasion of colorectal cancer HCT116 and RKO cells and the two show synergy. The mechanism is the likelihood that the combination down-regulates the expression of PI3K and Akt.
Abstract：ObjectiveTo investigate the effect of Wudan pill on the polarization of macrophages in the rat model of endometriosis （EMT） with cold congeal and blood stasis syndrome based on p38 mitogen-activated protein kinases （p38 MAPK）.MethodFemale SD rats with regular motility cycles were selected and randomly divided into sham-operated group， model group， Wudan pill high， medium， and low-dose groups （2.4， 1.2， and 0.6 g⋅kg-1）， Chinese patent medicine group， and western medicine group by the random number table method. The method of ice water bath + autologous endometrial transplantation was used to establish the rat model of EMT with cold congeal and blood stasis， and the rats were executed after 4 weeks of continuous drug administration to collect materials. Expression levels of serum tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-4， and transforming growth factor-β （TGF-β） were determined by enzyme-linked immunosorbent assay （ELISA） to assess inflammation. The real-time quantitative polymerase chain reaction （Real-time PCR） was performed to determine the inducible nitric oxide synthase （iNOS）， TNF-α， arginase 1 （Arg1）， and human mannose receptor （CD206） transcriptional levels to evaluate macrophage polarization. Western blot （WB） and immunofluorescence （IF） assays were used to determine the protein expression levels of iNOS and Arg1 to corroborate macrophage polarization. WB and Real-time PCR were used to determine the protein expression levels of the p38 MAPK pathway.ResultAs compared with sham-operated group， the levels of serum TNF-α， IL-1β， TGF-β， and IL-4 of rats in the model group were significantly higher （P<0.05）. In the model group， the protein levels of iNOS， TNF-α， p-p38 MAPK， and phosphorylated-extracellular signal-regulated kinase （p-ERK） in endothelial tissues were significantly higher， the mRNA levels of iNOS， TNF-α， MAPK， and ERK were significantly higher， and the mRNA and protein expression levels of Arg1 and CD206 were significantly lower （P<0.05， P<0.01）. The number of iNOS positive cells in endothelial tissues was significantly increased， and the number of Arg1 positive cells in endothelial tissues in the model group was significantly decreased （P<0.05， P<0.01）. As compared with the model group， the expression of TNF-α， IL-1β， TGF-β， and IL-4 in each administration group was reduced to different degrees， which was especially significant in the Wudan pill high and medium-dose groups and the western medicine group （P<0.05）. The protein expression levels of iNOS， TNF-α， p-p38 MAPK， and p-ERK in endometrial tissues of rats in the Wudan pill high and medium-dose groups， the Chinese patent medicine group， and the western medicine group were significantly lower， the mRNA expression levels of iNOS， TNF-α， MAPK， and ERK were significantly lower， and the protein expression levels of Arg1 and CD206 were significantly higher （P<0.05， P<0.01）. The number of iNOS positive cells in endometrial tissues of rats was significantly decreased in the Wudan pill high and medium-dose groups， the Chinese patent medicine group， and the western medicine group， whereas the number of Arg1 positive cells was increased （P<0.05， P<0.01）. The low， medium， and high doses of Wudan pill were dose-dependent， and the efficacy of the Wudan pill high-dose group was similar to that of the western medicine group.ConclusionWudan pill reduces the inflammatory response in rat model of EMT with cold congeal and blood stasis syndrome and decreases expression levels of TNF-α， IL-1β， IL-4， and TGF-β， thereby prompting polarization of macrophages from M1 to M2 type. The mechanism is presumedly related to p38 MAPK signaling pathway.
Keywords：endometriosis;cold congeal and blood stasis;inflammatory factors;macrophage polarization;p38 mitogen activated protein kinases
Abstract：ObjectiveTo investigate the effect of supplemented Buyang Huanwutang on kidney tissue， nuclear factor-κB （NF-κB） pathway， and fibrosis factors in diabetic kidney disease （DKD） mice.MethodA total of 24 db/db mice （11-12 weeks old） were randomized into the model group （equivalent volume of distilled water， once/day， 8 weeks）， supplemented Buyang Huanwutang group （16.0 g·kg-1， once/day， 8 weeks）， and irbesartan group （13.5 mg·kg-1， once/day， 8 weeks） after adaptive feeding for 1 week and positive urinary protein monitoring， with 8 in each group. Another 8 db/m mice （11-12 weeks old） were included in the normal group （equivalent volume of distilled water， once/day， 8 weeks）. Then samples were collected， and the levels of fasting blood glucose （FBG）， total cholesterol （TC）， triglyceride （TG）， blood urea nitrogen （BUN）， serum creatinine （SCr）， and urinary microalbumin （mALB） were detected. The mRNA expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and monocyte chemoattractant protein-1 （MCP-1） was determined by real-time polymerase chain reaction （Real-time PCR）. The expression of nuclear factor-κB （NF-κB）， NF-κB inhibitor α （IκBα）， phosphorylated IκBα （p-IκBα）， transforming growth factor-β1 （TGF-β1）， α-smooth muscle actin （α-SMA）， and fibronectin （FN） in kidney tissue was measured by Western blot. The expression of NF-κB in renal tissue was detected by immunofluorescence. The pathological changes of kidney were observed under light microscope.ResultCompared with the normal group， the model group showed glomerular hypertrophy， increase in extracellular matrix， thickening of basement membrane， small cystic lumen， interstitial inflammatory cell infiltration， and some interstitial fibrosis （P<0.01）. Moreover， the model group had higher content of FBG， mALB， TC， TG， BUN， and SCr （P<0.01）， higher expression of inflammatory factors TNF-α， IL-1β， and MCP-1， and fibrosis-related proteins TGF-β1， α-SMA， and FN （P<0.01）， and stronger activation of NF-κB pathway in renal tissue （P<0.01） than the normal group. Compared with the model group， supplemented Buyang Huanwutang alleviated the pathological injury in kidney （P<0.01）， decreased the content of mALB， TC， and TG， the content of BUN and SCr （P<0.01）， and the content of TNF-α， IL-1β， and MCP-1 （P<0.05， P<0.01）， and inhibited the activation of NF-κB pathway and the expression of fibrosis factors in renal tissue （P<0.05， P<0.01）， but had no significant effect on blood glucose level.ConclusionBy inhibiting NF-κB pathway and the expression of fibrosis factors in renal tissue， supplemented Buyang Huanwutang can exert anti-inflammatory and anti-fibrosis effect and alleviate the pathological damage in kidney tissue， thereby protecting the kidney.
Abstract：ObjectiveTo investigate the effect of capsaicin on cognitive dysfunction in rats with cerebral ischemia-reperfusion and its possible mechanism.MethodTwelve SD male rats were randomly selected as a sham operation group， and the remaining rats were sutured to replicate the model of middle cerebral artery occlusion （MCAO）. The successfully modeled rats were divided into a model group， a SB203580 ［p38 mitogen-activated protein kinase （p38 MAPK） inhibitor， 1 mg·kg-1］ group， capsaicin low- and high-dose （50， 100 mg·kg-1） groups ， and anisomycin （p38 MAPK agonist， 2 mg·kg-1） + capsaicin （100 mg·kg-1） group， with 12 rats in each group. After reperfusion and administration， the rats were scored for neurological deficits. Morris water maze and new object recognition experiments were used to test the learning and cognitive abilities of rats. The hematoxylin-eosin （HE） staining was used to observe the pathological changes in the hippocampus of the brain tissue. Immunofluorescence method was used to detect the activation of microglia in the hippocampus. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and prostaglandin E2 （PGE2） inflammatory factors in the brain tissue. Western blot was used to determine the protein expression levels of transient receptor potential vanillin subfamily 1 （TRPV1）， p38 MAPK， p-p38 MAPK， and cyclooxygenase-2 （COX-2） in the hippocampal tissue.ResultAs compared with the sham group， the neurological deficit score， escape latency， the number of Iba-1 positive microglia in the hippocampal CA1 area， the IL-1β， TNF-α， and PGE2 levels in the brain tissue， and the p-p38 MAPK/p38 MAPK and COX-2 expression in the hippocampus tissue was significantly increased in the model group （P<0.01）. In the model group， the number of crossing the platform position， the novel object discrimination index （DI）， and the TRPV1 expression in the hippocampus tissue was significantly reduced （P<0.01）， the number of hippocampal nerve cells was reduced， and a large number of inflammatory cells infiltrated. As compared with the model group， the neurological deficit score， escape latency， the number of Iba-1 positive microglia in the hippocampal CA1 area， the IL-1β， TNF-α， and PGE2 levels in the hippocampus tissue， and the p-p38 MAPK/p38 MAPK and COX-2 expression in the hippocampus tissue were significantly reduced in the capsaicin low-dose and high-dose groups （P<0.05，P<0.01）. In the capsaicin low-dose and high-dose groups， the number of crossing the platform position， the DI， and the TRPV1 expression in the hippocampus tissue were significantly increased （P<0.05，P<0.01）， a small amount of inflammatory cells were infiltrated， and the number of nerve cells was significantly increased. The use of anisomycin， an activator of p38 MAPK， increased the expression of COX-2， and significantly weakened the inhibitory effect of capsaicin on the activation of microglia.ConclusionCapsaicin has a protective effect on the cognitive function of rats with cerebral ischemia-reperfusion， and its mechanism may be related to the inhibition of the activation of p38 MAPK/COX-2 signaling pathway， thereby inhibiting the excessive activation of microglia.
Keywords：capsaicin;cerebral ischemia-reperfusion;cognitive dysfunction;p38 mitogen-activated protein kinase/cyclooxygenase 2 signaling pathway
Abstract：ObjectiveTo evaluate the clinical efficacy of Niuhuang Chengqitang in the treatment of acute gastrointestinal injury （AGI） in patients with severe traumatic brain injury （sTBI） and observe the effect on serum motilin （MTL）， diamine oxidase （DAO）， and neuron-specific enolase （NSE）.MethodBy the prospective randomized controlled trial （RCT） method， 86 sTBI inpatients with AGI were randomly divided into an observation group（43 cases）and a control group （43 cases）. All patients were given conventional western medicine treatment， including actively targeting the primary diseases and combined diseases， supply of enteral nutrition as soon as possible， and gastrointestinal agents， while those in the observation group received additional Niuhuang Chengqitang by nasal administration. The treatment course of both groups was 7 days. The clinical efficacy of the two groups was compared before treatment and 7 days after treatment. Gastrointestinal failure （GIF） score and intra-abdominal pressure （IAP） of the two groups were recorded， and serum MTL， DAO and NSE levels were detected before treatment， 3 days and 7 days after treatment. The Glasgow Coma Scale （GCS） score， duration of mechanical ventilation， length of ICU stay， and 28-day mortality were recorded.ResultDuring the treatment and observation， 4 cases fell off and 82 cases were finally enrolled， including 41 in the observation group and 41 in the control group. After treatment， the total effective rate in the observation group was 80.49%（33/41）， higher than 56.10%（23/41） in the control group （χ2=9.137，P<0.05）. GIF and IAP scores in the observation group were significantly improved after 3 and 7 days of treatment （P<0.01）， and the observation group was superior to the control group after 7 days of treatment （P<0.05）. The serum levels of MTL， DAO， and NSE in the two groups were improved after 3 and 7 days of treatment （P<0.05， P<0.01）. The levels of MTL and NSE in the observation group were better than those in the control group after 3 days of treatment （P<0.05， P<0.01）， and the levels of MTL， DAO， and NSE in the observation group were superior to those in the control group after 7 days of treatment （P<0.05， P<0.01）. The 28-day GCS score in the observation group was higher than that in the control group （P<0.01）， and the duration of mechanical ventilation and ICU stay in the observation group were significantly shorter than those in the control group （P<0.05）. There was no significant difference in 28-day mortality between the two groups. No adverse reactions related to Niuhuang Chengqitang occurred in the observation group.ConclusionNiuhuang Chengqitang can effectively improve the gastrointestinal function， regulate MTL， DAO， and NSE levels， shorten the ICU stay， and promote the recovery of sTBI patients with AGI.
Abstract：ObjectiveTo evaluate the clinical efficacy of modified Zhibai Dihuangwan on children （female） with idiopathic precocious puberty （ICPP） and to explore the mechanism.MethodA total of 78 children with ICPP who were treated in Henan Children's Hospital from July 2019 to July 2020 were randomized into the observation group and control group by sealed envelope randomization with 39 in either group. The observation group was given modified Zhibai Dihuangwan （1 dose/day， oral， divided into two times） and the control group Zhibai Dihuangtang （1 dose/day， oral， divided into two times orally）. The treatment lasted 3 months for both groups. The changes of mammary nucleus length， maximum follicular diameter， uterus， and ovarian volume before and after treatment in the two groups were observed， and serum luteinizing hormone （LH） level， follicle-stimulating hormone （FSH） level， prolactin （PRL） level， and estrogen （E2） level were detected in the two groups. At the cellular level， the secretion of gonadotropin releasing hormone （GnRH） and mRNA expression of precocious puberty-related genes Kiss-1 and G protein-coupled receptor 54 （GPR54） were measured.ResultLevels of serum sex hormones of the two groups were significantly decreased after treatment as compared with those before treatment （P<0.01）. After treatment， the length of the mammary nucleus and the maximum follicular diameter in the observation group were smaller than those in the control group （P<0.01）. The total clinical effective rate in the observation group was higher than that in the control group （P<0.01）. At the cellular level， the expression levels of GnRH， Kiss-1 and GPR54 mRNA in GT1-7 cells treated with modified Zhibai Dihuangwan were lower than those treated with traditional Zhibai Dihuangtang.ConclusionModified Zhibai Dihuangwan can effectively inhibit levels of serum sex hormones in children with ICPP and delay the development of mammary nucleus and ovary in children with precocious puberty. It is more effective than traditional Zhibai Dihuangtang， which is worthy of clinical promotion. The mechanism is the likelihood that it inhibits the expression of Kiss-1 and GPR54 mRNA.
Abstract：ObjectiveTo observe the clinical effect of modified Qilang prescription on the treatment of drug-dependent constipation of Qi-Yin deficiency type.MethodIn a randomized double-blind clinical trial， 160 patients with drug-dependent constipation were randomly divided into two groups， namely， the treatment group （80 cases） and the control group （80 cases）. The treatment group was treated with modified Qilang prescription and lactulose oral liquid simulator， while the control group was treated with lactulose oral liquid and the simulator of modified Qilang prescription. The course of treatment of both groups was 8 weeks. The scores of main syndromes of constipation， traditional Chinese medicine （TCM） syndromes， patient assessment of constipation symptom （PAC-SYM） scale， and patient assessment of constipation quality of life （PAC-QOL） scale before and after treatment were recorded. The patients were followed up at the 2nd and 4th week after drug withdrawal.ResultAfter treatment， the full analysis set （FAS） analysis showed that the total effective rate was 91.14% （72/79） in the treatment group and 73.33% （55/75） in the control group. The treatment group had a higher total effective rate than the control group （Z=-6.62， P<0.01）. The per-protocol set （PPS） analysis showed that the total effective rate was 93.51% （72/77） in the treatment group and 76.39% （55/72） in the control group. The treatment group had a higher total effective rate than the control group （Z=-6.77， P<0.05）. After treatment， the FAS and PPS analysis showed that the main syndrome scores of constipation were lower in both groups than those before treatment （P<0.05）. Except the syndromes of falling， overexertion defecation， and distension， the scores of other syndromes in the treatment group were lower than those in the control group （P<0.05）. The scores of TCM primary syndromes in the two groups were lower than those before treatment （P<0.05）， and the scores of the treatment group were significantly lower than those of the control group （P<0.05）. The scores of TCM secondary syndromes in the treatment group were significantly lower than those before treatment （P<0.05）. The syndrome scores of abdominal distension， fatigue， and lack of sleep in the treatment group were significantly lower than those in the control group （P<0.05）. The scores of PAC-SYM scale were improved in both groups after treatment （P<0.05）， and the treatment group was superior to the control group in improving fecal syndromes and abdominal syndromes. The scores of PAC-QOL scale were improved in both groups after treatment， and the treatment group was superior to the control group in improving patients’ physiological function， social psychology， anxiety， and satisfaction. After drug withdrawal， the recurrence rate was lower in the treatment group at the 2nd（χ2 =5.65，P<0.05） and 4th week （χ2 =12.37，P<0.01）.Conclusionmodified Qilang prescription is effective in the treatment of drug-dependent constipation， and its clinical effect is better than that of lactulose oral liquid. The curative effect of modified Qilang prescription is stable and lasting without obvious adverse reactions.
Keywords：modified Qilang prescription;drug-dependent constipation;lactulose;quality of life;clinical study
Abstract：ObjectiveTo investigate the clinical efficacy of negative pressure sealing drainage combined with compound Huangbai liquid on diabetic foot and effects on serum inflammatory factors.MethodA total of 168 patients with diabetic foot treated in Rizhao Hospital of Traditional Chinese Medicine from January 2019 to December 2021 were enrolled and randomly divided into a control group （84 cases） and an experimental group （84 cases）. All patients received basic treatment such as blood glucose control and anti-infection. The patients in the control group were treated with negative pressure drainage combined with normal saline，while those in the experimental group were treated with negative pressure drainage combined with compound Huangbai liquid. The ulcer areas，Visual Analogue Scale （VAS） scores，ankle-brachial index （ABI），CT angiography of the lower extremities （diameter of the dorsalis pedis artery，average blood velocity，and blood flow），and serum inflammatory factors［C-reactive protein （CRP），interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α）］ of the two group were compared before and after treatment，and the quality of life，satisfaction，and clinical efficacy of patients in the two groups were assessed after treatment.ResultCompared with the conditions before treatment，the two groups showed reduced ulcer areas，VAS scores，mean blood flow rate of dorsal foot artery，CRP，IL-6，and TNF-α levels （P<0.05），increased ABI，dorsum arterial blood flow，and lumen diameter（P<0.05），and improved quality of life （P<0.05）. Compared with the control group after treatment，the experimental group showed superior improvement in the ulcer area，VAS score，ABI，mean blood flow rate of dorsal foot artery，blood flow，and serum inflammatory factors（P<0.05），better quality of life，and higher satisfaction of treatment effect （P<0.05）. Under different treatment protocols，the total clinical effective rate of the experimental group was 92.86%（78/84），higher than 71.43%（60/84） in the control group （χ2=13.070，P<0.05）.ConclusionNegative pressure sealing drainage combined with compound Huangbai liquid for the treatment of diabetic foot can effectively relieve the clinical symptoms in patients，reduce ulcer area and VAS score，improve the blood circulation in dorsalis pedis artery，and decrease the levels of CRP，IL-6，and TNF-α，thereby inhibiting the occurrence of inflammatory reaction.
Abstract：ObjectiveBased on sequential metabolism and ultra-high performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）， to identify the prototype components and metabolites of Tongfengding capsules in rat plasma.MethodAn ACQUITY UPLC BEH Shield RP18 column （2.1 mm×100 mm， 1.7 µm） was used for gradient elution with 0.1% formic acid aqueous solution （A）-acetonitrile （B） as the mobile phase （0-1 min， 5%B； 1-2.4 min， 5%-10%B； 2.4-13.5 min， 10%-32%B； 13.5-18.5 min， 32%-90%B； 18.5-19 min， 90%-5%B； 19-21 min， 5%B） at a flow rate of 0.3 mL·min-1， injection volume of 2 μL and column temperature at 35 ℃. The heated electrospray ionization （HESI） was applied under positive and negative ion modes， and the scanning range was m/z 100-1 500. By comparing the differences between the administered plasma and the blank plasma， the prototype components and metabolites in intestinal metabolism sample and liver metabolism sample prepared by intestinal perfusion with parallel blood collection， and comprehensive metabolism sample prepared by intragastric administration method were identified.ResultA total of 76， 53， 74 chemical components were detected in the intestinal metabolism sample， the liver metabolism sample and the comprehensive metabolism sample. A total of 100 components were identified from these different plasma samples， including 64 prototype components （34 alkaloids， 12 terpenoids， 9 organic acids， 6 flavonoids and 3 other components） and 36 metabolites. The main metabolic reactions involved in the formation of metabolites were glucuronidation， deglucosylation， dehydrogenation and hydroxylation.ConclusionThe chemical components of Tongfengding capsules can undergo a series of metabolic reactions in the intestine and liver， and a large number of metabolites are generated， among which alkaloids may be the leading component group for efficacy， which can lay the foundation for systematic elucidating the material basis of Tongfengding capsules in vivo.
Keywords：Tongfengding capsules;sequential metabolism;ultra-high performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）;hydroxylation;glucuronidation;alkaloids;terpenoids
Abstract：ObjectiveTo construct the rat model of high-normal blood pressure with excessive phlegm-dampness syndrome and analyze the serum metabolites and explain the metabolic characteristics of the model rats.MethodThe model rats of high-normal blood pressure with excessive phlegm-dampness syndrome were fed with high-fat diet and intraperitoneal injected with 7.625 mg·kg-1 of Nω-nitro-L-arginine （L-NNA）， while rats in the normal group were fed chow diet and injected with the same amount of normal saline. The model was evaluated from the aspects of the general state， body weight， blood pressure and serum biochemical indexes， such as triglyceride （TG）， total cholesterol （TC）， high density lipoprotein cholesterol （HDL-C）， low density lipoprotein cholesterol （LDL-C）， blood glucose， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px）. Liquid chromatography-mass spectrometry was used to analyze the changes of metabolites in rats serum. The conditions were as follows：mobile phase of 0.05% formic acid aqueous solution （A）-0.05% formic acid acetonitrile solution （B） for gradient elution （0-3 min， 2%B； 3-9 min， 2%-40%B； 9-18 min， 40%-98%B）， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 80-1 000. Univariate and multivariate statistical analysis were used to screen the differential metabolites between groups which were used to metabolic pathways enrichment analysis.ResultCompared with the normal group， the rats in the model group had the characteristics of darker fur， lazy movement and loose stools， the body weight， blood glucose and contents of TG， TC， LDL-C， MDA increased significantly （P<0.05， P<0.01） and GSH-Px level decreased significantly （P<0.01）， the blood pressure remained stable at about 160/88 mmHg （1 mmHg≈0.133 kPa）. A total of 115 differential metabolites were screened in the positive and negative ion modes， of which 45 were up-regulated and 70 were down-regulated ［variable importance in the projection （VIP） value>1， P<0.05 and fold change （FC）≥2］， including indoleacetaldehyde， 5-hydroxytryptamine， lysophosphatidylcholine， phosphatidylcholine， xanthine and so on， which regulated blood pressure mainly through glycerophospholipid metabolism， linoleic acid metabolism， tryptophan metabolism and arachidonic acid metabolic pathways.ConclusionFeeding with high-fat diet and intraperitoneal injection of L-NNA can successfully construct the rat model of high-normal blood pressure with excessive phlegm-dampness syndrome， and indoleacetaldehyde， 5-hydroxytryptamine， lysophosphatidylcholine， phosphatidylcholine and xanthine can be used as characteristic serum differential metabolites of the rat model.
Abstract：ObjectiveIn this study， the two different origins of Lycii Cortex in the 2020 edition of Chinese Pharmacopoeia were determined to analyze their chemical consistency by comparing their main chemical composition.MethodThirty representative batches of Lycii Cortex were collected， content determination and fingerprint analysis methods were established by ultra performance liquid chromatography-photodiode array detector （UPLC-PDA） combining with multivariate statistical analysis to evaluate the similarities and differences between two origins of Lycii Cortex. Respectively by the mobile phase of acetonitrile （A）-0.15% trifluoroacetic acid aqueous solution （B） and the mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-4 min， 5%-12%A； 4-8 min， 12%A； 8-12 min， 12%-14%A； 12-15 min， 14%-30%A； 15-17 min， 30%-40%A； 17-18 min， 40%-90%A）， and the detection wavelength was set at 280 nm.ResultThis established content determination and fingerprint methods had good precision， stability and repeatability. The similarities of 30 batches of Lycii Cortex were above 0.90 by comparing with the control fingerprint， and the eight common peaks in fingerprints of Lycii Cortex from Lycium barbarum and L. chinense were all phenolic amides， which were kukoamine B， N-（4，9，13-triazatridecan-1-yl）-3，4-dihydroxybenzenepropanamide， feruloylputrescine， N1，N5-bis （dihydrocaffeoyl） spermidine or N5，N10-bis （dihydrocaffeoyl） spermidine， N5-caffeoyl-N10-dihydrocaffeoylspermidine， N5-dihydrocaffeoyl-N10-caffeoylspermidine， N1，N5-bis （caffeoyl） spermidine and lyciumin A. Among them， the content ranges of kukoamine B in Lycii Cortex from L. chinense and L. barbarum were 1.22%-8.18%， 2.52%-12.24%， respectively.ConclusionThe established UPLC analysis method can be used for the content determination and fingerprint analysis of Lycii Cortex. This study indicates that chemical contour of Lycii Cortex from L. barbarum and L. chinense are similar， there are no significant differences in kukoamine B content， and they have consistency in the chemical composition.
Abstract：ObjectiveTo clarify the accumulation and distribution characteristics of dry matter and mineral elements in Artemisia argyi var. argyi cv. Qiai， and to provide technical support for the high yield of and efficient utilization of nutrients in this medicinal species.MethodTwo cultivars of this species， Qiqing 1 and Qihuang 1 were selected， and the composition of dry matter in different organs， the content， accumulation， and distribution of mineral elements in each organ of the two cultivars， and the dynamic changes of volatile oil content and index components eucalyptol and borneol in leaves of the two cultivars were monitored at different growth stages.ResultThe period from February to March marked the early growth stage of Qiai， and the dry matter was mainly distributed in the leaves. It accelerated the growth in April， and the period from April to mid-June witnessed the vigorous vegetative growth of Qiai， during which the dry matter was mainly found in the stems and leaves. It began the reproductive growth from late June and the dry matter was mainly distributed in the stems. In the flowering stage in August， no dry matter accumulation occurred. As for the volatile oil， the content was high （> 1.10%） at the vigorous vegetative growth stage and peaked on June 14 （1.33% in Qiqing 1， and 1.23% in Qihuang 1）. The relative mass fraction of eucalyptol was the maximum at the vegetative growth stage （8.67% in Qiqing 1， and 13.07% in Qihuang 1）. The relative mass fraction of borneol peaked at the early growth stage （2.63% in Qiqing 1， and 5.94% in Qihuang 1）. The content of nitrogen， phosphorus， potassium， and zinc in leaves was in significantly positive correlation with the content of volatile oil and the relative content of eucalyptol and borneol. The content of macroelements nitrogen， phosphorus， potassium， and calcium and trace elements iron and zinc peaked at the early growth stage， and the content was the highest in stem and leaf. The content of macroelement magnesium and trace elements manganese and copper was the highest at vegetative growth stage when the content of other elements decreased and the nutrients were gradually transferred to the buds， flowers and other organs. In the whole growth period， the distribution of potassium， calcium， and zinc was in the order of leaf > stem > root， and the distribution of nitrogen， phosphorus， copper， magnesium， and manganese followed the order of leaf > root > stem. The distribution of iron was in the order of root > leaf > stem. There was a significantly positive correlation between the total amount of dry matter and the absorption of nutrients in 'Qiai'. The absorption of macroelements by Qiai was in the order of potassium > nitrogen > calcium > phosphorus > magnesium， and the ratio of absorbed elements was about 2.66∶2.51∶0.6∶0.11∶0.04. The absorption of trace elements followed the order of manganese > iron > zinc > copper， and the ratio of absorbed elements was about 0.25∶0.17∶0.05∶0.04. In terms of the production of medicinal materials， 'Qiai' needed about 4.11 kg potassium， 3.58 kg nitrogen， 0.91 kg phosphorus， 0.18 kg calcium， 0.06 kg magnesium， about 6.64 g manganese， 2.56 g iron， 1.30 g zinc， and 0.92 g copper to produce 100 kg medicinal materials.ConclusionEnough organic fertilizer and phosphorus and potassium fertilizers should be applied as base fertilizers for Qiai. The vegetative growth stage （April-June） marks the high accumulation of dry matter and large demand of nutrients， during which topdressing should be conducted timely and early， especially nitrogen fertilizer， and appropriate amount of micro-element fertilizer should be added. Qiai needs a large amount of calcium and magnesium fertilizers from the mid-vegetative growth stage， and they should be applied in time in the late stage to ensure the vegetative growth of the plants for seeds and the quality of the medicinal material of Qiai.
Abstract：ObjectiveTo study the accumulation and absorption of dry matter and nutrients in Curcuma phaeocaulis from Sichuan， the origin of Curcumae Radix （tuber） and Curcumae Rhizoma （rhizome）， to explore the growth and development laws and nutrient demand characteristics of the medicinal species， and thus to lay a theoretical basis for rational fertilization.MethodThe plant growth indexes， dry matter accumulation， and nutrient content of C. phaeocaulis at different growth stages in Sichuan were measured and analyzed.ResultThis medicinal species featured the dominant growth of aboveground leaves and stems before October and growth of underground part （particularly the rhizomes and tubers） from October. During the whole growth period， the accumulation of nitrogen， phosphorus， potassium， calcium， magnesium， iron， manganese， zinc， and copper per plant was 2 450.31， 907.09， 3 171.18， 625.94， 493.38， 14.53， 2.24 ， 2.93， 0.46 mg， respectively， with the order of potassium > nitrogen > phosphorus > calcium > magnesium > iron > zinc > manganese > copper.ConclusionThe species needs sufficient potassium and nitrogen， appropriate amount of phosphorus， calcium， and magnesium， a small amount of iron， and very little zinc， manganese， and copper for growth， and potassium is particularly important. Nitrogen， phosphorus， and potassium fertilizers supply macroelements and nitrogen fertilizer should be supplemented at seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome. Phosphorus and potassium fertilizers should be applied at seedling stage， tuber expansion stage， secondary expansion stage of rhizome， and dry matter accumulation stage of tuber and rhizome. At seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome， a variety of medium and trace elements-containing foliar fertilizers should be used.
Keywords：Curcuma phaeocaulis;dry matter;nutrients;growth and development law;nutrient demand characteristics
Abstract：Bronchiectasis is a refractory chronic airway disease characterized by chronic cough and phlegm production. Airway mucus hypersecretion is one of the important causative factors for acute exacerbation of bronchiectasis. Thus， reducing airway mucus secretion during the stable phase of bronchiectasis is the key to the treatment. Phlegm-dispelling therapy is commonly used in modern medicine， which， however， fails to radically reduce mucus secretion. At the moment， it is difficult and urgent to find an intervention that can improve the quality of life of patients and reduce the recurrence of disease during the stable phase of bronchiectasis. Chinese medicine shows remarkable efficacy in reducing airway mucus secretion. Nonetheless， there is a lack of research on the airway mucus hypersecretion of bronchiectasis， and no registered research outcome is available. Traditional Chinese medicine boasts a lot of experience in the treatment of phlegm. It combines clearing and tonifying methods and identifies Yin and Yang and internal organs， thus being advantageous in the treatment of airway mucus hypersecretion of bronchiectasis. Therefore， it is an important topic to improve the effectiveness of Chinese medicine on airway mucus hypersecretion of bronchiectasis by focusing on the "generation"， "transformation" and "excretion" of phlegm.
Keywords：traditional Chinese medicine;bronchiectasis;airway mucus hypersecretion;clearing method;tonifying method
Abstract：Glaucoma is the first irreversible blinding eye disease in the world. The current animal models of glaucoma are mainly divided into ocular hypertension type and non-ocular hypertension optic nerve damage type according to the presence or absence of ocular hypertension. Based on the clinical diagnostic criteria and symptoms of glaucoma in traditional Chinese medicine （TCM） and western medicine，this study summarized and analyzed the classification，modeling methods，advantages and disadvantages of common animal models of glaucoma，and the degree of agreement between TCM and western medicine. Among the existing animal models of glaucoma，those induced by laser photocoagulation，anterior chamber injection，episcleral vein cauterization，and circumcorneal and scleral limbus suture，and transgenic models have a high degree of agreement with western medicine diagnosis and main eye syndromes in TCM， suggesting that the models available can ideally show the clinical characteristics of glaucoma in western medicine. However， the TCM etiology and pathogenesis are rarely considered in the existing animal models of glaucoma， and the information on four examinations and TCM syndrome characteristics have not been observed. Moreover， research on the combination of diseases and syndromes in animal models of glaucoma has not been carried out. Therefore，it is necessary to explore and establish an animal model of glaucoma with the combination of TCM and western medicine that is in line with clinical practice. This study aimed to explore unified diagnostic criteria and evaluation system of TCM and western medicine for animal models of glaucoma with syndrome-disease combination and considered how to reflect the characteristics of TCM syndromes based on glaucoma with liver depression syndrome to establish a glaucoma model with syndrome-disease combination in animals suitable for clinical practice with the characteristics of TCM and western medicine， thereby promoting higher-quality research on glaucoma.
Keywords：glaucoma;animal model;clinical disease;traditional Chinese and Western medicine
Abstract：Chicoric acid， a hydroxycinnamic acid with the molecular formula C22H18O12， is an important active ingredient in Taraxacum mongolicum， Echinacea purpurea， Cichorium intybus and other natural plants， and it helps plants protect themselves from insects and infection from viruses， bacteria， fungi， and nematodes. Modern pharmacological research suggests that chicoric acid has significant bioactivities such as anti-inflammatory， antioxidant， immune-regulating， antibacterial， antiviral and anti-tumor properties. The first reported bioactivity of chicoric acid is its inhibitory effect on human immunodeficiency virus （HIV）. With the development and application of molecular biology and related technologies， the inhibitory activities of chicoric acid on other viruses as well as its mechanism of action have been frequently reported. Another study indicates that chicoric acid has significant inhibitory effects on different pathogenic bacteria. This paper summarized the research progress on the antiviral and antibacterial effects of chicoric acid through a comprehensive review of relevant literature in China and abroad in the past 20 years. Studies have shown that chicoric acid has significant inhibitory activities against various viruses such as HIV， hepatitis B virus （HBV）， respiratory syncytial virus （RSV）， and herpes simplex virus （HSV）， as well as different pathogenic bacteria such as Staphylococcus aureus， Yersinia enterocolitica and Mycobacterium tuberculosis. This study is expected to provide references for in-depth research on chicoric acid against pathogenic microorganisms and antiviral and antibacterial study of traditional Chinese medicine.
Abstract：Boiling is a common processing method of Chinese medicine. Based on the book of Summary of Processing Methods Data of Traditional Chinese Medicine in Past Dynasties， the authors consulted herbal books in all ages， combined with modern processing laws and regulations in various provinces and cities， the boiling methods and Chinese medicine varieties in ancient and modern times， judgment method of the endpoint of processing， as well as the study on boiling methods of representative Chinese medicines were compiled and analyzed. After sorting， it was found that the application of boiling methods began in the Han dynasty， enriched and developed in the Northern and Southern dynasties， Tang， Song and Yuan dynasties， and reached its heyday in the Ming and Qing dynasties. However， the number of modern boiling varieties decreased and mainly focused on toxic Chinese medicines or those that need to change or moderate their medicinal properties， indicating the development of boiling methods entered a stable period. The varieties of excipients used in the modern age mainly considered factors such as convenience of use and easy access， and the boiling degree， time and times were commonly used to judge the endpoint of boiling process. The main purposes of using boiling method for Chinese medicines were to remove impurities， remove non-medicinal parts， change or moderate the medicinal properties， and eliminate or reduce adverse reactions， which can provide a reference for carrying out the common research of boiling method for Chinese medicines.
Keywords：processing of Chinese medicines;boiling method;historical evolution;decoction pieces;endpoint of processing
Abstract：Among the gastrointestinal cancers， gastric cancer （GC） has high incidence， which is the third leading cause of cancer-related death worldwide after lung cancer and liver cancer. The survival rate of patients with GC is low and the prognosis is poor， posing a threat to the quality of life and leading to unmanageable financial burden of patients. As evidenced， it is crucial for the prevention and treatment of GC to block or reverse precancerous lesions of gastric cancer （PLGC） at the early stage. At the moment， the pathogenesis of PLGC is still unclear. The classical nuclear factor kappa B （NF-κB） pathway is widely involved in the inflammatory response， proliferation， apoptosis， angiogenesis and other processes of tumor cells， interfering with a variety of signaling molecular activities related to tumorigenesis. The dysregulation of NF-κB signaling pathway will accelerate the transformation of PLGC to GC. Accumulating evidence shows that NF-κB is closely related to PLGC and GC and they influence each other. Therapeutic drugs targeting NF-κB pathway have been widely studied， especially compound Chinese medicine preparations， active components of Chinese medicinals， and NF-κB inhibitors. By inhibiting the chronic inflammatory response of gastric mucosa， angiogenesis， and the proliferation， invasion， and migration of atypical cells， promoting apoptosis， and anti-tumor mechanism， they block the carcinogenesis of gastric epithelial cells， thereby accurately targeting the PLGC and preventing and treating GC. This article reviews the mechanism of NF-κB signaling pathway in the pathogenesis of PLGC and the treatment with Chinese and western medicine based on NF-κB pathway， which is expected to provide theoretical support and scientific basis for clinical treatment and prevention of PLGC.
Abstract：The diverse nervous system diseases （NSDs） are common complex refractory systemic diseases in clinical settings， which are manifested as trigeminal neuralgia， facial paralysis， neuromyelitis optica， multiple sclerosis， stroke， vascular dementia， encephalitis B， Parkinson's disease， epilepsy， Alzheimer's disease， amyotrophic lateral sclerosis， and diabetic peripheral neuropathy. Amid the accelerating population aging， the morbidity and prevalence of NSDs have been on the rise， posing a threat to quality of life of patients and bringing a heavy burden to society and individuals. This is also a challenge to the clinical prevention and treatment of the diseases. At the moment， NSDs are mainly treated with western medicine which is effective. However， in the long-term use and progression of the diseases， there are some potential risks such as adverse drug reactions， easy recurrence after withdrawal， drug dependence， and drug resistance. From the holistic view， traditional Chinese medicine （TCM） excels in treating diseases based on syndrome differentiation. According to the etiology， location， and pathogenesis of diseases in TCM， the overall therapy is applied， shows obvious advantages in alleviating disease symptoms， delaying disease progression， and improving the quality of life. TCM has a long history and categorizes NSDs into encephalopathy. According to clinical practice and theories， toxin is an important pathogen of NSDs in TCM. To be specific， the external toxins are insect and beast toxins， food toxin， toxin of six excesses， and epidemic toxin， and the internal toxins are phlegm toxin， dampness toxin， stasis toxin， fire toxin， and turbid toxin. Accumulation of all pathogens will lead to diseases with multiple syndromes. In the case of sudden attack of toxin， the disease is characterized by rapid onset. In the instance of fierce toxin， disease progresses rapidly. If the toxin is dependent， the disease symptoms are diverse. If the toxin is stubborn and frequently occurs， disease recurrence is common. In the case that toxin damages the collaterals， the healthy Qi is impaired. Controlling physiological function of the body， nervous system corresponds to the house of brain in TCM， which is the extraordinary organ， the house of intelligence， and the seat of mental activities that governs the whole body. In the case of toxin damaging the brain collaterals， cerebrospinal injury and orifice obstruction will occur， followed by disharmony of Qi and blood， Yin-yang imbalance， vital activity injury， disorder of primordial spirit， and dysfunction. The treatment principle should be detoxification and eliminating pathogen， and different diseases should be treated with the same method. At the moment， many doctors and scholars have focused on the prevention and treatment of NSDs based on the toxin pathogen. This paper summarizes the research on the clinical application of toxin pathogen causing disease in NSDs， including the peripheral nerve disease， demyelinating disease of the central nervous system， cerebrovascular disease， infectious disease of central nervous system， movement disorders， epilepsy， nervous system degenerative disease and nervous system complications of internal medicine disease， which is expected to provide some ideas and methods for the prevention and treatment of NSDs with TCM.
Keywords：toxin pathogen causing disease;nervous system;encephalopathy in TCM;clinical application
Abstract：Diabetic wounds are slow to heal， which poses a challenge to the medical field. Being vulnerable to infection， they are a major cause of amputation and even death and thus are costly. Chronic inflammation is an important culprit of the lingering diabetic wounds. NOD-like receptor protein 3 （NLRP3）， apoptosis associated speck-like protein （ASC）， and aspartate-specific proteasezymogen procaspase-1 （pro-Caspase-1）， constitute an intracellular protein complex called the NLRP3 inflammasome. Activated NLRP3 inflammasome can induce the release of pro-inflammatory factors interleukin（IL）-1β and IL-18 and participate in a variety of inflammatory responses. The activation of the NLRP3 inflammasome is associated with several inflammatory diseases. It has been concluded that many factors such as microcirculation disorder of diabetic wounds， accumulation of advanced glycation end products， oxidative stress injury， and long-term infiltration of macrophages can influence NLRP3 inflammasome， which induce persistent inflammation of the wounds. Therefore， solutions to the diabetic wound， such as targeting the NLRP3 inflammasome， reducing its hyperactivation， and inhibiting its overexpression， have emerged. Based on the correlation between the pathological changes of diabetic wounds and NLRP3 inflammasome， this article summarized the research on the methods of reducing NLRP3 inflammasome expression to promote the healing of diabetic wounds， such as regulating diabetic wound oxidative stress， balancing neutrophil extracellular traps （NETs） / NLRP3 inflammasome axis， inducing macrophage M2 polarization， reducing the production of advanced glycation end products， and enhancing autophagy. Moreover， the mechanisms of active constituents of Chinese medicine and compound Chinese medicine prescriptions against NLRP3 inflammasome activation were analyzed. Thereby， this paper is expected to provide new targets for diabetic wound healing and a reference for research the mechanism of Chinese medicine in anti-inflammation and promoting healing.
Keywords：diabetic wound healing;NOD-like receptor protein 3 （NLRP3） inflammasome;Chinese medicine
Abstract：Diabetic kidney diseases （DKD） is one of the main microvascular complications of diabetes. According to the available studies， the initial factor of DKD is glomerular injury. However， in recent years， more and more studies have confirmed that renal tubular injury plays a key role in the development of DKD. Taking renal tubules and their interstitium as therapeutic targets has become a new idea for the treatment of DKD. Traditional Chinese medicine （TCM） can treat renal tubular injury in DKD via multiple pathways， targets， and links， demonstrating significant therapeutic effect and slight side effects. According to the etiology and pathogenesis of DKD， doctors differentiate different syndromes such as deficiency of both Qi and Yin， deficiency of spleen and kidney Yang， and Qi deficiency and blood stasis and then adopt the therapies of nourishing kidney and strengthening essence， clearing heat and eliminating dampness， tonifying Qi and nourishing yin and so on， which have shown significant effect in the treatment of renal tubular injury in DKD. This paper summarizes the experimental studies about the treatment of renal tubular injury in DKD by TCM compound prescriptions， as well as the effective parts， extracts， and active ingredient of Chinese herbal medicines. Chinese herbal medicines and their derivates can regulate Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB）， nuclear factor erythroid 2-related factor 2 （Nrf2）/antioxidant response element （ARE）， transforming growth factor-β1 （TGF-β1）/Smads and other signaling pathways to treat renal tubular injury from resisting oxidative stress， inhibiting apoptosis of renal tubular epithelial cells， and blocking renal tubular epithelial-mesenchymal transformation. With this review， we hope to provide a theoretical basis for the development and clinical application of new drugs for renal tubular injury in DKD.
Keywords：traditional Chinese medicine;diabetic kidney disease;renal tubular injury;clinical research;fundamental research
Abstract：Ulcerative colitis（UC）is a disease characterized by chronic persistent inflammation of the colorectal mucosa. Its complex pathological mechanism is related to immune inflammation and enhanced apoptotic activity. The Janus kinase（JAK）/signal transducer and activator of transcription（STAT）is an important regulatory pathway in the body's physiological function, which can regulate the release of intestinal pro-inflammatory factors and induce apoptosis, resulting in colon tissue damage. In the condition of UC, the biological activities and expression levels of JAK and STAT increased, and the tissue inflammatory response and apoptosis rate increased, which led to the destruction of intestinal mucosal tissues. At present, in the treatment of UC, glucocorticoids and immunosuppressants are mainly employed to reduce intestinal inflammation. Although they can block the progress of UC to some extent, the adverse reactions are severe. A large number of studies have shown that traditional Chinese medicine（TCM） has significant advantages in the prevention and treatment of UC and can significantly reduce the recurrence rate of this disease. In recent years, plenty of studies have been carried out to explore the role of TCM in the treatment of UC by regulating the JAK/STAT pathway. The results have shown that the JAK/STAT pathway is the key target pathway of TCM in the treatment of UC. Based on the etiology and pathogenesis of deficiency and excess, TCM regulates the JAK/STAT pathway by clearing heat, drying dampness, cooling and activating blood, invigorating the spleen, warming the kidney, and performing both tonification and elimination to maintain the balance between pro-inflammatory factors and anti-inflammatory factors, weaken colonic inflammatory response, inhibit apoptosis, and play a role in the treatment of UC. The present study analyzed the mechanism and effect of TCM in intervening in UC by targeting the JAK/STAT signaling pathway and summarized the molecular mechanisms of different cytokines such as interleukin-6（IL-6）, IL-10, IL-23, microRNA（miRNA）-146a, and suppressors of cytokine signaling 2/3（SOCS2/3） on many family subtypes of the JAK/STAT signaling pathway to facilitate the comprehensive understanding of researchers on the mechanism of TCM on the JAK/STAT pathway in UC, which is expected to provide a theoretical basis for the treatment of UC and further drug development.
Keywords：traditional Chinese medicine;Janus kinase（JAK）;signal transducer and activator of transcription（STAT）;ulcerativecolitis（UC）;signaling pathway