Abstract：ObjectiveTo observe the effect of classical prescription Gegen Qinliantang（GGQLT） on inflammatory factors and key targets in the inflammatory pathways mediated by lipopolysaccharide in KKAy mice and explore its mechanism in improving spontaneous type 2 diabetes mellitus （T2DM）.MethodSixty-five SPF KKAy mice with spontaneous T2DM and 13 C57BL/6J mice （control） were selected in the barrier system and fed on a high-fat diet. The model was properly induced in 44 mice in the context of random blood glucose exceeding or equal to 13.9 mmol·L-1. Then the mice were assigned into a normal group （20 mL∙kg-1 normal saline）， a model group （20 mL∙kg-1 normal saline）， an acarbose group （3.9 mg∙kg-1）， and high- and low-dose GGQLT groups （1.82 and 0.45 g∙kg-1）， with 11 mice in each group. The mice in each group were treated correspondingly by gavage for eight weeks， once per day. Blood glucose and body weight were systematically evaluated. Twelve hours after the last administration， blood samples were collected from the eyes， and the serum and muscle and liver tissues were extracted. The levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and glucose transporter type 4 （GluT4） were detected by semi-quantitative enzyme-linked immunosorbent assay （ELISA）. The protein expression of IκB kinase β （IKKβ） and nuclear factor-κB （NF-κB） in muscle tissues and Toll-like receptor 4 （TLR4） in liver tissues was detected by Western blot.ResultCompared with the normal group， the model group showed increased body weight and blood glucose （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced body weight and blood glucose （P<0.05， P<0.01）. As revealed by ELISA results， compared with the normal group， the model group showed increased levels of TNF-α and IL-6 （P<0.01） and deceased GluT4 level （P<0.05）. Compared with the model group， the groups with drug treatment showed reduced levels of TNF-α and IL-6 （P<0.05， P<0.01）， and the acarbose group and the high-dose GGQLT group showed increased GluT4 level （P<0.05， P<0.01）. As displayed by Western blot results， compared with the normal group， the model group showed increased protein expression of IKKβ， NF-κB， and TLR4 （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced protein expression of IKKβ， NF-κB， and TLR4 （P<0.05， P<0.01）.ConclusionGGQLT can inhibit the inflammatory cascade effect and improve T2DM by down-regulating the levels of key inflammatory factors in the TLR4 pathway， inhibiting their activation， and increasing the translocation and activity of GluT4 on the basis of the regulation of intestinal flora.
Abstract：ObjectiveTo observe the effect of Liuwei Dihuangtang （LWDHT） on depression-like behaviors of rats with diabetes mellitus and depression （DD） and explore its mechanism.MethodThe diabetes mellitus （DM） model was induced by the high-fat diet and tail vein injection of low-dose streptozotocin （STZ） in 50 male Sprague-Dawley rats of SPF grade. Then the DD model was induced by chronic unpredictable mild stress （CUMS） for 28 days in DM rats. Fifty DD rats were randomly divided into model group， fluoxetine group （10 mg·kg-1·d-1）， and low-， medium-， and high-dose LWDHT groups （3.375， 6.75， 13.5 g·kg-1·d-1）， with 10 rats in each group. Another 10 healthy rats were assigned into a control group and received normal saline by gavage. After four weeks of drug intervention， the forced swimming assay was carried out to assess the depression-like behaviors of rats. The expression levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， and interleukin-10 （IL-10） in the anterior cingulate cortex （ACC） were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence was used to detect the expression of myelin basic protein （MBP） in ACC and the co-localization of ionized calcium-binding adapter molecule 1 （Iba1） with intracellular microtubule-associated protein 1 light chain 3 （LC3）. The protein expression levels of MBP， myelin proteolipid protein （PLP）， myelin oligodendrocyte glycoprotein （MOG）， Beclin-1， LC3， p62， and microglia （MG） phenotypic protein-related inducible nitric oxide synthase （iNOS）， and arginase 1 （Arg1） were detected by Western blot.ResultCompared with the control group， the model group showed shortened swimming time and prolonged immobility time （P<0.01）. Compared with the model group， the medium- and high-dose LWDHT groups showed reduced immobility time （P<0.05， P<0.01）. Compared with the control group， the model group showed decreased protein expression of MBP， PLP， and MOG in the ACC region （P<0.01）. Compared with the model group， the fluoxetine group and the medium- and high-dose LWDHT groups showed up-regulated protein expression of MBP， PLP， and MOG （P<0.05， P<0.01）. Compared with the control group， the model group showed decreased MBP fluorescence intensity in the ACC region （P<0.01）. Compared with the model group， the fluoxetine group and the medium- and high-dose LWDHT groups showed increased MBP fluorescence intensity in the ACC region （P<0.05， P<0.01）. Compared with the control group， the model group showed increased expression of iNOS （P<0.01） and slightly increased Arg1 protein expression. Compared with the model group， the medium- and high-dose LWDHT groups and the fluoxetine group showed down-regulated iNOS expression and up-regulated Arg1 protein expression （P<0.05， P<0.01）， but there was no significant difference between the fluoxetine group and the medium-，high-dose LWDHT groups. Compared with the control group， the model group showed increased expression levels of proinflammatory factors IL-1β and TNF-α in the ACC region （P<0.01） and slightly increased expression levels of anti-inflammatory factors IL-4 and IL-10. Compared with the model group， the fluoxetine group， and the medium- and high-dose LWDHT groups showed down-regulated expression of IL-1β and TNF-α （P<0.05， P<0.01） and up-regulated expression of IL-4 and IL-10 （P<0.05， P<0.01）. Compared with the control group， the model group showed reduced expression levels of Beclin-1 and LC3Ⅱ （P<0.01） and increased expression level of p62 （P<0.01）. Compared with the model group， the fluoxetine group and the medium- and high-dose LWDHT groups showed up-regulated Beclin-1 and LC3Ⅱ expression （P<0.01） and down-regulated p62 expression （P<0.01）. Compared with the control group， the model group showed decreased LC3+Iba1+ cells in the ACC region （P<0.01）. Compared with the model group， the fluoxetine group and the medium- and high-dose LWDHT groups showed increased LC3+Iba1+ cells （P<0.05， P<0.01）.ConclusionLWDHT can alleviate the depression-like behaviors in DD rats presumedly by promoting MG autophagy， regulating MG phenotypic changes， and increasing MG clearance of myelin sheath fragments. Meanwhile， MG phenotypic transformation also inhibits ACC inflammation in DD rats， improves the local microenvironment of oligodendrocyte proliferation and differentiation， and ultimately promotes the repair and remyelination of damaged myelin sheath.
Abstract：ObjectiveThis study investigated the mechanism of Wenjingtang in the prevention and treatment of endometriosis （EMT） from the perspective of regulating hypoxia stress and mitochondrial function.MethodPrimary human endometrial stromal cells （ESCs） form ectopic endometrial tissues were isolated and cultured， the cells were divided into control group （Control）， 5% control serum group （5% KBXQ）， 10% control serum group （10% KBXQ）， 5% Wenjingtang serum group （5% WJTXQ） and 10% Wenjingtang serum group （10% WJTXQ）. ESCs in different groups were detected for proliferation by methyl thiazolyl tetrazolium （MTT） assay， mRNA and protein expression of hypoxia inducible factor-1α （HIF-1α） by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot analysis， mitochondrial ultrastructure by transmission electron microscope， mitochondrial function ［mitochondrial membrane potential， mitochondrial membrane potential（MMP） and cytochrome C（Cyt C） content］ and apoptosis （cell membrane permeability， nuclear fluorescence intensity， nuclear size and cell counts） by high content screening （HCS） assay， apoptosis rate by flow cytometry， and proteins of B-cell lymphoma-2 （Bcl-2） associated X （Bax）， Bcl-2 and cleaved cysteinyl aspartate specific proteinase-3 （cleaved Caspase-3） by Western blot.ResultCompared with Control group， the 5% KBXQ and 10% KBXQ groups showed increased cell viability （P<0.01）， there was no significant change in HIF-1α mRNA and protein expression， transmission electron microscopy showed that the mitochondrial cristae were obvious and the inner and outer membranes of mitochondria were clear， HCS multichannel fluorescence staining showed that there were no significant changes in the expression of MMP， Cyt C and cell membrane permeability， and the nuclei showed uniform light staining， there were no significant changes in apoptosis rate， cleaved Caspase-3 protein expression and Bax/Bcl-2 ratio. Compared with Control group and corresponding concentration KBXQ group， the 5% WJTXQ and 10% WJTXQ group showed decreased cell viability （P<0.01） and HIF-1α mRNA and protein levels （P<0.05，P<0.01）， the ultrastructure of mitochondria was destroyed， some mitochondria were swollen and the cristae were blurred， moreover， decreased MMP and up-regulated Cyt C release （P<0.05，P<0.01）， increased cell membrane permeability （P<0.01）， and apoptosis characteristics included nuclear pyknosis， DNA agglutination in nucleus and decrease of cell numbers were observed （P<0.05，P<0.01）， increased apoptosis rate （P<0.01）， which was consistent with the results of HCS analysis， and up-regulated expression levels of cleaved Caspase-3 protein and Bax/Bcl-2 ratio （P<0.05，P<0.01）.ConclusionIn conclusion， the results suggest that Wenjingtang can improve hypoxia stress via down-regulating HIF-1α expression in ectopic ESCs， and inhibit cell proliferation， reduce mitochondrial biological activity and induce apoptosis， which might be the internal mechanism of Wenjingtang in preventing and treating EMT.
Abstract：ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Chifengtang on middle cerebral artery embolism（MCAO） rat model.MethodThe 90 SPF male rats were randomly divided into sham operation group， model group， high， medium and low dose groups of Huangqi Chifengtang （8.10，4.05，2.025 g·kg-1） and positive drug group （Naoxintong 0.32 g·kg-1）. MCAO rat model was established and intervened with Huangqi Chifengtang， and the neurological scores of each group were scored. The area of cerebral infarction was calculated by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Serum interleukin-6 （IL-6） and interleukin-1β（IL-1β） were detected by enzyme-linked immunosorbent assay （ELISA），The contents of matrix metalloproteinase-9（MMP-9）， vascular endothelial growth factor（VEGF） and vascular endothelial growth factor receptor 2 （VEGFR2）， the pathological changes of brain tissue in each group were observed by hematoxylin eosin （HE） staining. Western blot was used to detect zonule atresia protein-1（ZO-1）， tight junction protein-5（Claudin-5） and P-glycoprotein （P-gp） and multidrug resistance protein 1（MRP1）.ResultCompared with the sham operation group， the neurological function score and cerebral infarction rate of the model group were significantly increased（P<0.01）， and the levels of IL-6， IL-1β and MMP-9 in serum were significantly increased（P<0.01）， the levels of ZO-1 and Claudin-5 protein expression decreased significantly（P<0.01）， and the levels of P-gp and MRP1 protein expression increased significantly（P<0.01）. Compared with the model group， the neurological function score of rats in each administration group decreased significantly at 14 days （P<0.05，P<0.01）， the pathological changes of brain tissue effectively improved， the rate of cerebral infarction significantly reduced （P<0.01）， and the expression level of IL-6， IL-1β and MMP-9 in serum decreased significantly （P<0.05，P<0.01）， the content of VEGFR2 increased significantly （P<0.01）， and the content of VEGF increased significantly in high， medium dose and positive drug groups （P<0.05，P<0.01）. Although it decreased in low dose group， there was no significant difference. The levels of ZO-1 and Claudin-5 protein expression in brain tissue of high dose group and positive drug group increased significantly （P<0.05，P<0.01）， the level of MRP1 and P-gp protein expression decreased significantly （P<0.05）.ConclusionHuangqi Chifengtang can play a therapeutic role in rats with cerebral infarction by improving the pathological changes of brain tissue， reducing inflammatory reaction， promote angiogenesis and regulating the function of blood-brain barrier（BBB）.
Keywords：Huangqi Chifengtang;cerebral infarction;inflammation;angiogenesis;blood brain barrier;tight junctional protein
Abstract：ObjectiveTo observe the effects of Wumeiwan-medicated serum on the proliferation， invasion， migration， and apoptosis of human pancreatic cancer SW1990 cells and explore the underlying mechanism.MethodThe Wumeiwan-medicated serum was prepared and the pancreatic cancer SW1990 cell line was cultured in vitro. The optimal time of Wumeiwan-medicated serum was selected for subsequent experiments by cell counting kit-8（CCK-8）. SW1990 cells were divided into a control group and low- （2%）， medium- （4%）， and high-dose （8%） Wumeiwan-medicated serum groups. The colony-forming， migration， and invasion abilities were detected by clonogenic assay， wound healing assay， and transwell migration assay. Flow cytometry was used to detect the effect of Wumeiwan-medicated serum on the apoptosis of pancreatic cancer SW1900 cells. Western blot was used to detect the expression levels of apoptosis-related proteins， such as B-cell lymphoma-2（Bcl-2）， Bcl-2-associated X protein （Bax）， cytochrome C （Cyt C）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）， cleaved cysteinyl aspartate-specific protease-9 （cleaved Caspase-9）， as well as phosphatidylinositol 3-kinase（PI3K）， phosphorylated PI3K（p-PI3K）， protein kinase B （Akt）， and phosphorylated Akt（p-Akt）in PI3K/Akt pathway in SW1990 cells.ResultCompared with blank group， Wumeiwan groups showed decreased absorbance （A） 72 h after drug intervention （P<0.01）. Compared with the low-dose group， the medium- and high-dose groups showed decreased A （P<0.01）. Compared with the medium-dose group， the high-dose group showed decreased A （P<0.01）. It indicates that Wumeiwan can inhibit SW1990 cell proliferation in a dose-dependent manner after 72 h， and the optimal action time is 72 h. Compared with the blank group， the Wumeiwan groups showed weakened invasion of SW1990 cells （P<0.01）， reduced colony-forming and migration abilities （P<0.05， P<0.01） in a dose-dependent manner， and increased total apoptosis rates （P<0.01）. The inducing effect of Wumeiwan on apoptosis increased with the increase in dosage. Compared with the blank group， the Wumeiwan groups showed decreased protein expression of Bcl-2 （P<0.01）， increased protein expression of cleaved Caspase-3， cleaved Caspase-9， Cyt C， and Bax （P<0.05， P<0.01） in a certain dose-effect relationship， reduced protein expression of p-PI3K and p-Akt （P<0.05， P<0.01） with the increase in dosage， and declining p-PI3K/PI3K and p-Akt/Akt （P<0.05， P<0.01） with the increase in dosage.ConclusionWumeiwan-medicated serum can significantly inhibit the malignant biological behaviors of pancreatic cancer SW1990 cells and induce apoptosis. The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway and down-regulation of protein phosphorylation level in the PI3K/Akt signaling pathway.
Keywords：Wumeiwan-medicated serum;pancreatic cancer;proliferation;invasion;metastasis;phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway
Abstract：ObjectiveTo explore the effects and related mechanisms of modified Shuyuwan on the decline of learning and memory in Alzheimer's disease （AD） mice.MethodForty 5-month-old SPF APP/PS1 mice were randomly divided into model group， Donepezil group， modified Shuyuwan group， modified Shuyuwan+ chloroquine （CQ） group， 10 mice in each group， the same background wild type C57BL/6J ten mice were set as the normal group. Among them， the modified Shuyuwan group was given the modified Shuyuwan decoction （10 g·kg-1）， the Donepezil group was given the Donepezil hydrochloride solution （0.45 mg·kg-1）， the modified Shuyuwan + CQ group was CQ （10 mg·kg-1） was injected intraperitoneally on the basis of the modified Shuyuwan group， and the normal group and the model group were given the same amount of normal saline intragastrically， once a day， for a total of 35 days. After the administration， Morris water maze experiment and new object recognition experiment to detect the spatial memory ability of mice. TdT-mediated dUTP Nick-End Labeling（TUNEL） staining to detect the apoptosis level of mouse hippocampal CA1 neurons， biochemical detection of reactive oxygen species （ROS） and superoxide in mouse hippocampal neurons dismutase （SOD） levels. transmission electron microscopy to observe the ultrastructure of neuronal mitochondria in the CA1 region of mouse hippocampus. Western blot to detect mouse hippocampal mitochondrial autophagy adaptor protein （p62） and microtubule-associated protein 1 light chain 3 Ⅱ （LC3Ⅱ）， PTEN-induced kinase 1 （PINK1）， E3 Ubiquitin Ligase（Parkin）protein expression level. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） detection of mouse hippocampal mitochondrial forkhead transcription factor O1 （FoxO1）， PINK1， Parkin mRNA expression level.ResultCompared with the normal group， the escape latency of the model group mice increased significantly， the number of crossing platforms and the retention time in the target quadrant decreased significantly， the relative resolution index decreased significantly， and the ability to recognize new objects was weakened （P<0.05）， neurons in the hippocampus CA1 area decreased. The number of dead cells increased significantly （P<0.05）， the level of ROS was significantly increased （P<0.01）， and the level of SOD was significantly decreased （P<0.01）， the morphology of hippocampal mitochondria was severely damaged， the expression of p62 and LC3Ⅱ proteins increased （P<0.01）， Parkin protein expression decreased （P<0.05）， and PINK1 protein expression increased （P<0.05）， FoxO1， PINK1， Parkin mRNA expressions all decreased （P<0.05）. Compared with the model group， the mice's escape latency was significantly shortened after the intervention of the modified Shuyuwan， the number of crossing platforms and the proportion of residence time in the target quadrant increased significantly， the relative resolution index increased significantly， and the ability to identify new objects was enhanced （P<0.05）. Apoptotic cells were significantly reduced （P<0.05）. ROS levels were significantly reduced （P<0.01）， and SOD levels were significantly increased （P<0.05， P<0.01）， mitochondrial morphology and various structures were significantly improved， p62， LC3Ⅱ protein expression decrease （P<0.05，P<0.01）， PINK1， Parkin protein expression increased （P<0.01）. FoxO1， PINK1， Parkin mRNA expression increased （P<0.05， P<0.01）. Compared with the modified Shuyuwan group， the evasion latency of mice in the modified Shuyuwan + CQ group increased significantly， the number of crossing platforms and the proportion of residence time in the target quadrant decreased， and the relative resolution index decreased （P<0.05）， the SOD level was significantly reduced （P<0.01）. The damage of mitochondrial morphology and structure increased again， the expression of p62 and LC3Ⅱ protein increased （P<0.05， P<0.01）， and the expression of PINK1 and Parkin decreased significantly（P<0.05， P<0.01）. FoxO1， PINK1， and Parkin mRNA expression was significantly reduced （P<0.05， P<0.01）.ConclusionModified Shuyuwan can effectively improve the oxidative stress damage and learning and memory ability of AD mice. The mechanism may be related to up-regulating the expression of FoxO1， PINK1， and Parkin factors， promoting mitochondrial autophagy， reducing oxidative stress， and protecting neuronal damage.
Abstract：ObjectiveTo explore the mechanism of coking death and apoptosis of A549 cells induced by Tingli Dazao Xiefeitang.MethodA549 cells were randomized into blank group， traditional Chinese medicine（TCM） low， medium， and high concentration groups， which were treated with 20， 40， 60 mg·L-1 Tingli Dazao Xiefeitang， and TCM low， medium， and high concentration groups， respectively， and blank group was treated with equal volume culture medium. After 48 h of treatment， cell migration was detected by scratch assay and cell apoptosis was detected by flow cytometry. The relative expression levels of cysteine aspartate protease-1（Caspase-1）， NOD-like receptor protein 3 （NLRP3）， dermoderin D （GSDMD）， Survivin protein and nuclear transcription factor -κB （NF-κB） pathway proteins were detected by Western blot. The levels of intracellular reactive oxygen species （ROS） were determined by DCFH-DA fluorescence probe， and the contents of tumor necrosis factor -β （TNF-β） and interleukin-1β （IL-1β） in supernatant were determined by enzyme-linked immunosorbent assay （ELISA）.ResultCompared with blank group， the scratch healing rate， apoptosis rate， relative expression of Survivin protein， Caspase-1， GSDMD， NLRP3， ROS and NF-κB phosphorylation levels were significantly increased in low， medium and high concentration groups. The contents of TNF-β and IL-1β in supernatant were significantly increased （P<0.05）. Compared with the low concentration group， the scratch healing rate， apoptosis rate， Survivin protein relative expression， Caspase-1， GSDMD， NLRP3 relative expression， ROS and NF-κB phosphorylation levels were significantly increased in the medium and high concentration groups. The contents of TNF-β and IL-1β in supernatant were significantly increased （P<0.05）. Compared with the TCM group， the scratch healing rate， apoptosis rate， Survivin protein relative expression， Caspase-1， GSDMD， NLRP3 relative expression， ROS and NF-κB phosphorylation levels were significantly increased in the high concentration group. The contents of TNF-β and IL-1β in supernatant were significantly increased （P<0.05）.ConclusionTingli Dazao Xiefeitang can improve NLRP3 protein expression， inhibit Survivin protein expression and promote apoptosis of A549 cells. At the same time， it can activate NF-κB pathway and ROS system， up-regulate the expression of Caspase-1 and GSDMD， mediate scortosis of A549 cells.
Abstract：ObjectiveTo explore the differences in the protective effects of five formulas for promoting blood circulation and removing blood stasis on the aortic endothelial cells of New Zealand rabbits with heart blood stasis syndrome.MethodEighty New Zealand rabbits were randomly divided into a normal group （n=10） and an experimental group （n=70）. The heart blood stasis syndrome model was induced by starvation combined with a high-fat diet and adrenaline in the rabbits of the experimental group. Subsequently， the model rabbits were randomly divided into a model group， a Xuefu Zhuyutang group （3.55 g·kg-1·d-1）， a Taohong Siwutang group （2.66 g·kg-1·d-1）， a Danshenyin group （1.962 g·kg-1·d-1）， a Huoluo Xiaolingdan group （2.80 g·kg-1·d-1）， a Shixiaosan group （0.56 g·kg-1·d-1）， and a c-Jun N-terminal kinase （JNK） inhibitor （SP600125， 5 μg·kg-1）group. The normal group and the model group received the same amount of distilled water. The rabbits in five Chinese medicine groups were treated correspondingly by gavage， and those in the SP600125 group were injected with 0.5 mL of SP600125-dimethyl sulfoxide diluent. After the treatment， the aorta was collected， and terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was used to detect the apoptosis of aortic endothelial cells. The enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）. Western blot was used to detect the protein expression of JNK， phosphorylated JNK （p-JNK）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cysteinyl aspartate-specific protease-9 （Caspase-9）， and cysteinyl aspartate-specific protease-3 （Caspase-3） in aortic tissues. Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA levels of JNK， Bcl-2， Bax， Caspase-9， and Caspase-3 in aortic tissues.ResultFive formulas could improve the apoptosis of aortic endothelial cells to varying degrees. To be specific， Xuefu Zhuyutang and Taohong Siwutang were optimal in efficacy， followed by Huoluo Xiaolingdan， Shixiaosan， and Danshenyin， and SP600125 was the worst （P<0.05， P<0.01）. Five formulas could reduce the content of TNF-α and IL-6 （P<0.05， P<0.01）， down-regulate the protein expression levels of JNK， p-JNK， Bax， Caspase-9， and Caspase-3 （P<0.05， P<0.01）， decrease the mRNA expression levels of JNK， Bax， Caspase-9， and Caspase-3 （P<0.05， P<0.01）， and up-regulate the protein and mRNA expression levels of Bcl-2 （P<0.05， P<0.01）.ConclusionFive formulas can all reduce the apoptosis of aortic endothelial cells in New Zealand rabbits with heart blood stasis syndrome with different efficacies. It may be related to the different effects of five formulas on the JNK signaling pathway.
Keywords：blood stasis syndrome;endothelial cell;c-Jun N-terminal kinase （JNK）;formula for promoting blood circulation and removing blood stasis
Abstract：ObjectiveTo observe the effects of Hedysarum polysaccharides（HPS）on the signaling pathways of B-cell lymphoma 2 （Bcl-2）， cysteinyl aspartate-specific protease 3 （Caspase-3）， and Bcl-2-associated X protein （Bax） in Schwann cells（SCs）cultured in high glucose，and explore the possible mechanism of HPS against diabetic peripheral neuropathy（DPN）.MethodFour SD suckling mice aged 5-7 days were randomly divided into a normal group，a high-glucose group，an HPS + high-glucose group，and an α-lipoic acid（α-LA）+ high-glucose group. SCs were extracted from the sciatic nerve and cultured in a 37 ℃，5% CO2 incubator. After the cells reached 80% confluence，Cell Counting Kit-8（CCK-8）was used to screen the experimental concentrations suitable for high glucose，HPS， and α-LA interventions. Western blot and Real-time polymerase chain reaction （Real-time PCR）were used to detect the protein and mRNA expression of Bcl-2，Bax，and Caspase-3. The apoptosis rate of SCs was detected by flow cytometry using Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI）.ResultAs revealed by Western blot and real-time PCR，compared with the normal group，the high-glucose group showed reduced protein and mRNA expression of Bcl-2 and increased protein and mRNA expression of Bax and Caspase-3（P<0.01）. Compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed increased protein and mRNA expression of Bcl-2 and decreased protein and mRNA expression of Bax and Caspase-3（P<0.01）. As displayed by the results of flow cytometry using Annexin V/PI， compared with the normal group，the high-glucose group showed increased apoptosis rate；compared with the high-glucose group，the HPS + high-glucose group and the α-LA + high-glucose group showed reduced apoptosis rate（P<0.01）.ConclusionHPS can alleviate the apoptotic response of SCs，and its mechanism may be related to the inhibition of the activation of the Bcl-2/Caspase-3 signaling pathway.
Keywords：Hedysarum polysaccharides;diabetic peripheral neuropathy;Schwann cells;apoptotic response;B-cell lymphoma 2 （Bcl-2）;cysteinyl aspartate-specific protease 3 （Caspase-3）;Bcl-2-associated X protein （Bax）
Abstract：ObjectiveTo investigate the inhibitory effect of Astragalus polysaccharide （APS） on epithelial-mesenchymal transition （EMT） induced by transforming growth factor-β1 （TGF-β1） in cisplatin （DDP）-resistant lung adenocarcinoma cell line A549/DDP cells transplanted into nude mice and the molecular mechanism in improving DDP resistance.MethodBALB/c nude mice were randomly divided into a blank group， a model group， a DDP group， and a combination group （APS combined with DDP）. A549/DDP cells were infected with TGF-β1-overexpressed lentiviral vector and the negative control. The infected cells were inoculated subcutaneously in nude mice. The A549/DDP cells with TGF-β1 gene overexpression were inoculated into all groups except the control group with negative TGF-β1 gene overexpression. The drug intervention was performed eight days after cell inoculation. The mice in the combination group received intragastric administration of APS （0.3 g·kg-1·d-1） and intraperitoneal injection of cisplatin （0.003 5 g·kg-1）， and those in the cisplatin group received intraperitoneal injection of cisplatin （0.003 5 g·kg-1）. After 32 days of cell inoculation， the nude mice were killed and the tumor tissues and lungs were collected. The tumor weight was recorded and the inhibition rate was calculated. The number of metastatic nodules of the lung tumor on the whole slide was counted under the microscope. Immunohistochemistry， Western blot， and real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and gene expression of EMT molecular markers α-catenin and N-cadherin， and tumor drug resistance markers human lung resistance protein （LRP）， multidrug resistance-associated protein （MRP）， and P-glycoprotein （P-gp） in the transplanted tumor.ResultCompared with the blank group， the model group showed increased tumor weight and metastatic nodules of the lung tumor （P<0.05）， decreased protein and mRNA expression of α-catenin （P<0.05）， and elevated protein and mRNA expression of N-cadherin， LRP， MRP， and P-gp （P<0.05）. Compared with the model group and the cisplatin group， the combination group showed reduced tumor weight and metastatic nodules of the lung tumor （P<0.05）， increased protein and mRNA expression of α-catenin （P<0.05）， and decreased protein and mRNA expression of N-cadherin， LRP， MRP， and P-gp （P<0.05）.ConclusionAPS can inhibit the growth and metastasis of the transplanted tumor of lung adenocarcinoma and improve cisplatin resistance， which may be related to the inhibition of EMT of tumor cells.
Abstract：ObjectiveTo study the effect of Longshengzhi capsule （LSZC） on high fat diet （HFD）-induced atherosclerosis （AS） in apolipoprotein E knockout （ApoE-/-） mice.MethodApoE-/- mice were fed with HFD for 8 weeks to induce AS. Then the mice were randomized into model group， simvastatin group （4 mg·kg-1）， high-dose LSZC group （1.6 g·kg-1）， medium-dose LSZC group （0.8 g·kg-1）， and low-dose LSZC group （0.4 g·kg-1）. C57BL/6J Mice with normal diet were used as the blank control. After 10 weeks， serum levels of triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， malondialdehyde （MDA）， superoxide dismutase （SOD）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） were detected. Hematoxylin-eosin （HE） and oil red O were used to detect aortic plaque in each group. The levels of CD34 and F4/80 in aorta were determined by immunohistochemistry （IHC）.ResultCompared with the blank control， the model group demonstrated obvious aortic plaque， a large amount of lipid accumulation， serious damage of aortic intima， increase in serum levels of TC， TG， LDL-C， HDL-C， MDA， IL-1β， and IL-6 （P<0.01）， decrease in SOD level （P<0.01）， and rise of the expression of CD34 and F4/80 （P<0.01）. Compared with the model group， LSZC of the three doses all decreased the serum levels of TG and LDL-C （P<0.05）， and the levels of IL-1β and IL-6 （P<0.05， P<0.01）， and the high-dose and medium-dose LSZC improved SOD level， decreased MDA content （P<0.05， P<0.01）， and reduced the expression of the CD34 and F4/80 in blood vessels （P<0.05， P<0.01）.ConclusionLSZC has certain intervention effect on the formation of aortic plaque in atherosclerosis ApoE-/- mice. The mechanism is that it reduces the levels of serum TG and LDL-C to lower blood lipid， decreases MDA level and improves SOD activity to inhibit lipid peroxidation， lowers the levels of IL-1β and IL-6 and down-regulates the expression of CD34 and F4/80 to protect blood vessels from inflammatory damage.
Keywords：Longshengzhi capsule;apolipoprotein E knockout （ApoE-/-） mice;atherosclerosis;aortic plaque;lipid accumulation
Abstract：ObjectiveTo observe the repair effect of Dahuanglingxian prescription （DHLX） on bile duct epithelial cells of rats. To explore whether its mechanism of action is to adjust the mutual binding of transforming growth factor -β （TGF-β） activated kinase 1（TAK1） and tumor necrosis factor receptor-associated factor 6 （TRAF6）， and regulate the activation of the nuclear transcription factor -κB （NF-κB）/mitogen-activated protein kinase （MAPK） signaling pathway.MethodThe 20 SD rats were randomly divided into normal group and DHLX group， 10 rats in each group， were given saline and DHLX （320 mg·kg-1·d-1） for 8 days， to prepare normal serum and DHLX serum. Biliary epithelial cells were extracted from normal SD rats and divided into 9 groups： Normal group， model group （20 mg·L-1）， LPS+DHLX group （20 mg·L-1+10% DHLX）， LPS+PDTC group （20 mg·L-1+200 μmol·L-1）， LPS+SB203580 group （20 mg·L-1+0.5 μmol·L-1）， LPS+PDTC+SB203580 group （20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1）， LPS+PDTC+DHLX group （20 mg·L-1+200 μmol·L-1+10% DHLX serum）， LPS+SB203580+DHLX group （20 mg·L-1+0.5 μmol·L-1+10% DHLX serum）， LPS+PDTC+SB203580 +DHLX group （20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1+10% DHLX serum）. The microscopic observation of morphological changes in each group of cells after drug intervention. Enzyme-linked immunosorbent assay（ELISA） was used to detect the expression of （IL）-1β and IL-6 in each group of cells. Western blot detected the expression levels of TAK1 and TRAF6 proteins in each group of cells， Co-IP detected the interaction between TAK1 and TRAF6， and further observed the distribution and co-localization of TAK1 and TRAF6 using Laser confocal microscope.ResultAfter the action of LPS， the cell synapses are reduced， the cell body becomes significantly rounded and smaller， but the cell morphology of each group tends to be normal after medication. Compared with normal group， the expression levels of IL-1β and IL-6 in model group were significantly increased （P<0.05）， while the expression level of TAK1 was decreased while the expression level of TRAF6 was increased （P<0.05）. The content of TAK1-TRAF6 protein complex showed a decreasing trend， and the two proteins co-located in the cytoplasm. Compared with model group， the expression levels of IL-1β and IL-6 in LPS+DHLX group were significantly decreased （P<0.05）， the expression level of TAK1 was increased and the expression level of TRAF6 was decreased （P<0.05）， the content of TAK1-TRAF6 protein complex was significantly increased （P<0.01）， and the two proteins were significantly co-located in cytoplasm. Compared with LPS+DHLX group， the expression levels of IL-1β and IL-6 in other groups were significantly decreased （P<0.05，P<0.01）. TAK1-TRAF6 protein complex content in each group was significantly decreased after pathway blocker intervention （P<0.05）， while TAK1-TRAF6 protein complex content in each group was significantly increased after pathway blocker combined with DHLX intervention （P<0.05）. Co-localization of the TAK1-TRAF6 in cytoplasm was not obvious.ConclusionIn the LPS-induced inflammatory response of bile duct cells， the binding of TAK1 and TRAF6 showed a weakening trend， but DHLX could reverse the phenomenon， we think the mechanism of action may be related to promoting the mutual binding of TAK1 and TARF6 to inhibit the activation of the NF-κB/MAPK signaling pathway.
Abstract：ObjectiveTo evaluate the effect of Wenshen Yangxue prescription （WSYX） on the outcome of in vitro fertilization-embryo transfer （IVF-ET） in poor ovarian responders and the safety.MethodA total of 116 eligible patients who were admitted to Beijing Obstetrics and Gynecology Hospital in June 2016-June 2019 were randomized into the experimental group and the control group with random number table method （58 in either group）. Conventional controlled ovarian hyperstimulation （COH） was directly implemented in the control group， while the experimental group was intervened with WSYX for 3 menstrual cycles before the COH. The pregnant patients were observed till childbirth and the non-pregnant patients for 12 months. Gonadotropins （Gn） dosage and the days of use were recorded. Serum levels of follicle-stimulating hormone （FSH）， basal FSH （bFSH）， basal luteinizing hormone （bLH）， and basal estrogen （bE2）， endometrial thickness， and antral follicle count （AFC） before and after treatment were measured. The serum levels of estrogen （E2）， progesterone （P）， and luteinizing hormone （LH） on the day of human chorionic gonadotropin （HCG） injection， and levels of FSH， LH， E2， testosterone （T）， and P in follicular fluid 36 h after HCG injection were determined. Number of fertilization， rate of fertilization， number of high-quality embryos， cycle cancellation rate， clinical pregnancy rate， and live birth rate were evaluated. The traditional Chinese medicine （TCM） syndrome scores before and after treatment were recorded. Liver and kidney functions were detected before and after treatment， and adverse reactions during treatment were recorded.ResultCompared with the control group， the experimental group showed the decrease in the days of Gn use and dosage of Gn （P<0.05）， endometrium thickening （P<0.05）， and increase in oocytes obtained， levels of E2 and LH on the day of HCG injection， and ovarian reserve function. Moreover， in follicular fluid 36 h after HCG injection， the reduction in level of FSH （P<0.05）， rise of levels of LH and E2 （P<0.05）， and insignificant changes in levels of T and P in the experimental group were observed as compared with those in the control group. In addition， larger number of fertilization， more available embryos， and higher rate of high-quality embryos were observed in the experimental group than in the control group （P<0.05）. The experimental group demonstrated improvement in quality of oocytes， decrease in cycle cancellation rate ， and increase in clinical pregnancy rate and live birth rate compared with the control group. The TCM syndrome score in experimental group was decreased after treatment compared with that before treatment （P<0.05）. No serious adverse reactions occurred in two groups during treatment and the safety indexes before and after treatment were all within the normal ranges.ConclusionWSYX can reduce the use duration and dosage of Gn in infertile patients receiving IVF-ET， improve the quality of oocytes， increase the rate of high-quality embryos， and improve the pregnancy outcome of IVF-ET patients.
Abstract：ObjectiveTo investigate the effect of Shunao Jieyu decoction on intestinal flora in patients with post-stroke depression.MethodSixty patients with post-stroke depression of Qi stagnation， blood stasis， and phlegm obstruction were selected and divided into a treatment group （30 cases， Shunao Jieyu decoction） and a control group （n=30， paroxetine hydrochloride tablets） according to the random number table. All patients were treated correspondingly for eight weeks. The scores of traditional Chinese medicine（TCM） syndrome， Hamilton rating scale for depression（HAMD）， National Institutes of Health stroke scale（NIHSS）， and activities of daily living（ADL）before and after treatment were compared between the two groups. High-throughput sequencing was used to analyze the diversity of fecal flora and the distribution of taxonomical levels in two groups before and after treatment.ResultThe post-treatment TCM syndrome score， HAMD score， and NIHSS score were lower than those before treatment in the same group （P<0.05）， while the post-treatment ADL score was higher than that before treatment （P<0.05）. Compared with the control group after treatment， the treatment group showed decreased TCM syndrome score （P<0.05）. No significant difference was observed in the HAMD score， NIHSS score and ADL score between the two groups after treatment. The total effective rate of the treatment group was 90% （27/30）， which was superior to 66.3% （19/30） of the control group （χ2=5.863， P<0.05）. After treatment， the average values of Chao1 index， Observed species index， Shannon index， Simpson index， and Pielou's evenness index of intestinal flora diversity in the treatment group increased without significant difference， while the average value of the Good's Coverage index remained unchanged in the same group. At the phylum level， the abundance of Bacteroidetes increased. At the family level， the abundance of Bacteroidaceae increased. At the genus level， the abundance of Bacteroidetes increased.ConclusionShunao Jieyu decoction can effectively improve the clinical TCM symptoms of patients with post-stroke depression， relieve neurological impairment， improve the ability of daily living， and change the diversity and abundance of the intestinal flora of patients at different taxonomic levels.
Abstract：ObjectiveTo investigate the effects of exogenous Fusarium oxysporum and Trichoderma viride on the diversity of soil fungal community and carbon metabolic function of cultivated Panax ginseng.MethodIllumina HiSeq 2500 high-throughput sequencing combined with Biolog-ECO was used to analyze the species diversity and functional diversity of soil fungal communities in P. ginseng soil under different exogenous treatments.ResultThe results of high-throughput sequencing showed that the number and species of microorganisms in the soil were significantly changed after exogenous microorganisms were added. The soil fungi with relative abundance greater than 1% included Mortierella sp.，Fusarium sp.，Humicola sp.，and Simplicillium sp. Mortierella sp. in each treatment group significantly increased. Humicola sp. and Simplicillium sp. could be induced to increase by exogenous addition of F. oxysporum，while T. viride at a high concentration could significantly inhibit the growth of F. oxysporum. As revealed by Biolog and principal component analysis （PCA），the average well color development （AWCD） in the high-dose T. viride group （MG） was significantly higher than that in the control group （QS）and the low-dose F. oxysporum group（LD）. The utilization abilities for amino acids，carboxylic acids，polymers， and amines were enhanced in the MG group，but the microbial metabolic activity was reduced in the high-dose F. oxysporum group （LG）. There was no significant increase in the utilization of phenolic acids by soil microorganisms in both groups.ConclusionExogenous addition of F. oxysporum can lead to the growth and reproduction of other pathogenic fungi. Exogenous addition of T. viride can enhance the soil fungal community structure and metabolic diversity，inhibit the proliferation of F. oxysporum，and improve the soil microbial environment of cultivated P. ginseng.
Keywords：Panax ginseng;cultivated Panax ginseng in the farmland;rhizosphere soil;high-throughput sequencing;Biolog-ECO
Abstract：ObjectiveTo optimize the existing genetic transformation system of Armillaria gallica to improve the transformation efficiency and lay a foundation for the follow-up research on Armillaria molecular marker-assisted breeding and gene function.MethodThe genetically transformed plasmid pH101-PAgGPD-GFP-TrpC was constructed，transformed into Escherichia coli，amplified， and cultured，and the plasmid was extracted. The extracted plasmid was transformed into four different agrobacteria LBA4404，EHA105，GV3101，and AGL-1，respectively. The transformed agrobacteria were used for impregnating A. gallica，and the agrobacteria with the highest conversion rate were screened out. Then the agrobacterium-mediated genetic transformation system of A. gallica was optimized from the type and concentration of antibiotics，co-culture time，concentration of bacterial solution， and impregnation method. The phenotype profiles of A. gallica under different conditions were observed using Synbiosis ProtoCol 3.ResultThe optimized genetic transformation conditions of A. gallica were as follows： the Agrobacterium strain of EHA105 at absorbance A600 nm=0.6， the co-culture time of 2 d， the infection mode of negative pressure impregnation for 10 min， the primary screening medium of PDA medium containing 400 mg·L-1 cefotaxime sodium and 10 mg·L-1 hygromycin，and the secondary screening medium of PDA medium containing 12 mg·L-1 hygromycin.ConclusionIn this study，the existing genetic transformation system of A. gallica was optimized，and there was a significant difference in the transformation rate before and after optimization （P<0.05）. After optimization，the transformation efficiency of A. gallica was about 4.33%，which was about eight times higher than that before optimization.
Abstract：ObjectiveTo explore the active ingredients， therapeutic targets， and relative signaling pathways of Tripterygium wilfordii in the treatment of triple negative breast cancer （TNBC） based on network pharmacology， and to verify the mechanism through in vitro cell model.MethodThe active ingredients of T. wilfordii were screened from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. The targets of TNBC were obtained from DisGeNET and GeneCards. Venny was used to identify the potential therapeutic targets of T. wilfordii against TNBC. Protein-protein interaction （PPI） network was constructed with String database. Gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were carried out with DAVID to predict the mechanisms of potential targets. The molecular docking between triptolide and key targets were performed with AutoDock Vina. The effect of triptolide （0， 5， 10， 20， 30， 40， 50， 60， 80 nmol·L-1） on the proliferation of MDA-MB-231 cells was determined through methyl thiazolyl tetrazolium （MTT） assay. The effect of triptolide （0， 12.5， 25， 50 nmol·L-1） on the apoptosis of MDA-MB-231 cells was detected with Hoechst 33342 staining. Western blot was performed to detect the effect of triptolide （0， 25， 50 nmol·L-1） on the expression levels of key targets.ResultT. wilfordii had 23 active ingredients related to 55 potential targets of TNBC. GO and KEGG enrichment revealed that the potential targets were associated with 103 biological processes， 15 cellular components， and 35 molecular functions， and were involved in 140 signaling pathways including atherosclerosis and apoptosis. The results of molecular docking demonstrated that triptolide could bind with the targets including threonine kinase 1 （Akt1）， vascular endothelial growth factor A （VEGFA）， cellular tumor antigen p53 （p53）， transcription factor AP-1 （JUN）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， mitogen-activated protein kinase 8 （MAPK8）， prostaglandin G/H synthase 2 （PTGS2）， and Caspase-3. According to the results of MTT assay， triptolide （20， 30， 40， 50， 60， 80 nmol·L-1） inhibited the proliferation of MDA-MB-231 cells compared with blank control （P<0.05， P<0.01）. Hoechst 33342 staining showed that triptolide （12， 25， 50 nmol·L-1） induced the apoptosis of MDA-MB-231 cells compared with black control （P<0.05， P<0.01）. Western blot showcased that 50 nmol·L-1 triptolide down-regulated the relative expression levels of p-Akt， TNF-α， and VEGFA， while 25 and 50 nmol·L-1 triptolide up-regulated the relative expression level of p53 in a dose-dependent manner compared with the blank control （P<0.05， P<0.01）.ConclusionT. wilfordii has multiple ingredients， targets， and pathways in the treatment of TNBC. It may regulate p53， VEGFA， TNF-α and other key targets to induce cell apoptosis and suppress angiogenesis and inflammatory response， which provides a scientific basis for the further investigation and clinical application of T. wilfordii.
Keywords：Tripterygium wilfordii;triple negative breast cancer;network pharmacology;molecular mechanism;experimental validation
Abstract：ObjectiveTo predict the pharmacodynamic basis and core target of Shengxiantang in the treatment of myasthenia gravis （MG） by network pharmacology and molecular docking and to further verify the molecular mechanism through animal experiment.MethodThe active components and potential targets of Shengxiantang were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and the disease-related targets from GeneCards and other databases. Then the common targets of the decoction and the disease were screened out， followed by the construction of protein-protein interaction （PPI） network， and Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of the common targets based on STRING database and Cytoscape 3.8.2. Afterward， Cytoscape 3.8.2 was employed to construct the disease-active component-target network. AutoDock and PyMOL were used for molecular docking of key components and hub genes. Finally， we used the Rα97-116 peptide to induce experimental autoimmune myasthenia gravis （EAMG） in rats and then verified the core target yielded in the docking with the model rats.ResultA total of 655 disease-related targets， 118 active components of the decoction， 21 common targets of the disease and the decoction， and 3 hub genes were screened out. The common targets were mainly involved in the GO terms of regulation of active oxygen metabolism， positive regulation of protein transport， and positive regulation of protein localization， and the KEGG pathways of toll-like receptor signaling pathway， phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， hypoxia inducible factor-1 （HIF-1） signaling pathway， and T cell receptor signaling pathway. The results of molecular docking showed that quercetin and Akt1 had the lowest and stable binding energy and interacted with each other through the amino acid residue LYS-30. Western blot demonstrated that Shengxiantang significantly inhibited the expression of p-Akt protein in the spleen of EAMG rats.ConclusionThe pharmacological mechanism of Shengxiantang in the treatment of MG may be that the main chemical components regulate the expression of the core protein Akt， and then may participate in and affect PI3K/Akt signaling pathways， laying a theoretical and experimental basis for further research.
Abstract：ObjectiveTo clarify the medication regularity of WU Zhao-dong，a famous chief physician in traditional Chinese medicine （TCM） of Jiangxi province， and investigate the potential mechanism of potential new prescriptions against chronic renal failure （CRF）.MethodThe outpatient prescriptions of WU Zhao-dong from July 2019 to July 2021 were collected. Data mining was carried out by using the Traditional Chinese Medicine Inheritance Auxiliary Platform （V 2.5） to analyze the medication frequency and drug association and obtain potential new prescriptions. The interaction between drug targets in new prescriptions was analyzed by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP），STRING，and Kyoto Encyclopedia of Genes and Genomes（KEGG）， followed by verification by molecular docking and experiments.ResultA total of 200 prescriptions were screened out， with 217 Chinese medicinal drugs involved， and eight new potential prescriptions were derived. To be specific， Prescription 1： Armeniacae Semen Amarum-Astragali Radix-Platycodonis Radix-Angelicae Sinensis Radix-Smilacis Glabrae Rhizoma-fried Atractylodis Macrocephalae Rhizoma， Prescription 2： Saposhnikoviae Radix-Schizonepetae Herba-Kochiae Fructus-Asteris Radix et Rhizoma-Menthae Haplocalycis Herba，Prescription 3：Armeniacae Semen Amarum-Asteris Radix et Rhizoma-Platycodonis Radix-Eriobotryae Folium-prepared Ephedrae Herba， Prescription 4：Perillae Caulis-Codonopsis Radix-Coptidis Rhizoma-Pseudostellariae Radix， Prescription 5：Ecliptae Herba-Astragali Radix Praeparata Cum Melle-Dryopteridis Crassirhizomatis Rhizoma-Rosae Laevigatae Fructus-Coicis Semen-Ligustri Lucidi Fructus， Prescription 6： Lycopi Herba-Lonice Raejaponicae Caulis-Trachelospermi Caulis et Folium-Alismatis Rhizoma， Prescription 7：Scutellariae Radix-Hirudo-Paeoniae Radix Rubra-Eriobotryae Folium-Glehniae Radix， Prescription 8：Glycyrrhizae Radix et Rhizoma-Scrophulariae Radix-Chrysanthemi Indici Flos-Smilacis Glabrae Rhizoma- Serissae Herba. In Prescription 1，18 main chemical components were screened out. Eighty targets of active components of Prescription 1 were predicted， and 37 potential targets for the treatment of CRF were obtained， including interleukin （IL）-6， vascular endothelial growth factor （VEGF）， cysteinyl aspartate-specific protease-3 （Caspase-3）， nitric oxide synthase 3 （NOS3）， and hypoxia-inducible factor-1 （HIF-1）. The KEGG pathways involved in the targets of Chinese medicinal drugs and disease mainly included the signaling pathways of lipid and atherosclerosis，NF-κB， Toll-like receptors， and HIF-1. Prescription 1 significantly decreased serum creatinine and urea nitrogen， and increased the content of NO and NOS3 in renal tissues of CRF rats.ConclusionPrescription 1 shows the multi-component and multi-target characteristics of action，and its mechanism may be related to its inhibition of renal fibrosis，anti-inflammation，improvement of intestinal microecology，and improvement of renal hypoxia and ischemia.
Abstract：ObjectiveTo screen the active antitumor components of Gupi Xiaoji decoction by network pharmacology and molecular docking based on the pyroptosis mediated by cysteinyl aspartate-specific protease 1 （Caspase-1） and explore its molecular mechanism in intervening in the pyroptosis of HepG2.2.15 cells through in vitro experiments.MethodThe compounds and targets of Gupi Xiaoji decoction were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） to obtain the corresponding gene symbols. The targets of Caspase-1 were collected from GeneCards，online mendelian inheritance in man（OMIM），PharmGKB，and TTD，and the compound-gene target regulatory network was constructed by Cytoscape. The protein-protein interaction（PPI） network was established and analyzed by STRING. The mechanism of the effective components of Gupi Xiaoji decoction on Caspase-1 was predicted by gene ontology（GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses. The molecular docking was verified with AutoDock Vina. The plasma medicated with Gupi Xiaoji Decoction was prepared and HepG2.2.15 cells were cultured in vitro. HepG2.2.15 cells were divided into a blank plasma group，a VX-765 group，a VX-765+medicated plasma group， and a medicated plasma group. After 48 hours of intervention with 15% medicated plasma， the expression and distribution of gasdermin D-N （GSDMD-N） on the surface of the cell membrane were detected by immunofluorescence staining. The release of lactic dehydrogenase （LDH）， interleukin（IL）-1β，and IL-18 in the cell supernatant was measured by enzyme-linked immunosorbent assay（ELISA） kits. The expression of Caspase-1 and GSDMD-N was measured by Western blot.ResultThe mitogen-activated protein kinase 14 （MAPK14），MAPK1，protein kinase B1 （Akt1）， MAPK8， V-Jun sarcoma virus oncogene homolog （JUN）， and TP53 screened by network pharmacology were the main targets. The compounds 7-hydroxy-5，8-dimethoxy-2-phenyl-chromone，wogonin，rhamnazin，moslosooflavone，isorhamnetin，7-O-methylisomucronulatol，formononetin，calycosin，luteolin，quercetin，kaempferol，β-sitosterol，and baicalein screened by network pharmacology were the main active components of Gupi Xiaoji decoction. Go enrichment analysis showed that multiple biological processes were involved， including responses to oxidative stress and metal ions，ubiquitin-like protein ligase binding，and phosphatase binding. KEGG pathway enrichment analysis showed MAPK pathway，nuclear factor（NF）-κB pathway，p53 pathway， and hypoxia-inducible factor-1（HIF-1） pathway were involved. Molecular docking showed that the targets had good binding with the components. In vitro experiments displayed that compared with the blank plasma group，the VX-765 group showed weakened GSDMD-N fluorescence signal，reduced release of LDH，IL-1β，and IL-18，and declining expression of Caspase-1 and GSDMD-N（P<0.01）， and the medicated plasma group showed increased GSDMD-N fluorescence signal， increased release of LDH，IL-1β，and IL-18，and up-regulated expression of Caspase-1 and GSDMD-N（P<0.01）.ConclusionGupi Xiaoji Decoction can induce the pyroptosis of HepG2.2.15 cells by regulating Caspase-1 through multiple targets and multiple pathways.
Abstract：ObjectiveTo evaluate the utility and mechanism of Huangqintang combined with carboplatin in chemotherapy of endometrial cancer by experiments as well as network pharmacology and molecular docking.MethodThe xenograft model of endometrial carcinoma was induced in BALB/c nude mice. When the tumor volume reached about 100 mm3，24 nude mice were randomly assigned into a model group， a Huangqintang group （3.5 g·kg-1），a carboplatin group （50 mg·kg-1），and a combination group （3.5 g·kg-1 Huangqintang + 50 mg·kg-1 carboplatin）， with six mice in each group. The mice in the model group received 200 μL of normal saline by gavage， twice a day. The volume of the tumor and the body weight of the mice were measured every two days. After drug intervention for 20 days， the blood of the mice was collected for renal function and blood routine tests. Then the nude mice were euthanized and the tumor was weighted. In combination with the experimental results，the underlying mechanism of Huangqintang combined carboplatin was predicted through network pharmacology and the binding sites of active components were predicted by molecular docking.ResultThe tumor inhibition rates of the Huangqintang group，the carboplatin group， and the combination group were 8.87%，50.33% （P<0.05），and 64.66% （P<0.01），respectively. Compared with the results in the model group，the body weight，leukocyte，erythrocyte， and hemoglobin in the carboplatin group decreased，and creatinine and uric acid increased （P<0.05）. Compared with the carboplatin group，the combination group showed increased body weight，leukocyte， and hemoglobin （P<0.05），and decreased creatinine and uric acid （P<0.05）. A total of 114 potential active components of Huangqintang involved 200 targets related to the side effects of carboplatin. The core genes involved were mainly heat shock protein 90AA1 （HSP90AA1），transcription factor c-Jun （JUN）， and mitogen-activated protein kinase （MAPK）. Molecular docking showed that baicalein and wogonin could form a stable protein complex with HSP90AA1， serving as potential active molecules. Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis revealed that it might be related to the regulation of tumor necrosis factor（TNF） signaling pathway，interleukin（IL）-17 signaling pathway， MAPK signaling pathway， and toll-like receptor pathway.ConclusionHuangqintang has no obvious inhibitory effect on endometrial cancer，and the tumor suppression effect is not significantly enhanced after combination with carboplatin，but Huangqintang can alleviate carboplatin-induced side effects. The mechanism may be related to the complex network of Chinese medicine.
Abstract：Traditional Chinese medicine （TCM） health technology assessment （HTA） provides scientific information and decision-making basis for decision-makers at all levels to choose TCM health technology reasonably. However， it is still in its infancy. In the future， it is necessary to strengthen top-level design， improve cognition and attention， enhance talent training and cooperation， and speed up the development of evaluation criteria in line with the characteristics of TCM， so as to promote the development of HTA in TCM. Therefore， it is imperative to create a method system and tools suitable for TCM HTA. Evidence and value impact on decisionmaking （EVIDEM） framework is developed by EVIDEM collaboration. It combines evidence and value to form a relatively complete decision-making framework system， which can provide methodological support for medical insurance reimbursement， clinical practice decision-making， drug selection and so on. The introduction of EVIDEM framework into the field of TCM has methodological feasibility and practical operability， but it is still necessary to further integrate and optimize the TCM characteristics. On the basis of theoretical research， pilot studies are carried out with Chinese patent medicine as the breakthrough point， forming a preliminary framework and implementation path for the research of TCM HTA， in order to promote healthcare decision-making and achievement transformation.
Keywords：traditional Chinese medicine;health technology assessment;evidence and value impact on decisionmaking （EVIDEM） framework;multiple criteria decision analysis;Chinese patent medicine;clinical comprehensive evaluation
Abstract：Xiao Xumingtang in The Catalogue of Famous Ancient Classics （The First Batch） issued by the National Administration of Traditional Chinese Medicine is derived from the Important Prescriptions Worth a Thousand Gold for Emergency （Bei Ji Qian Jin Yao Fang） written by SUN Si-miao in the Tang dynasty. The present study systematically explored the origin， development， historical evolution， and clinical application of Xiao Xumingtang. As revealed by the results， Xiao Xumingtang as well as its analogues are primary prescriptions indicated for apoplexy before the Tang and Song dynasties and serve as the benchmark for the treatment of apoplexy. After the Song dynasty， due to the changes in the understanding of the pathogenesis of apoplexy and the limitations of the understanding of Xiao Xumingtang， its clinical application to apoplexy gradually decreased. In modern times， it has been re-recognized and applied， during which its clinical applications have undergone great changes. Its clinical applications are extensive， involving a variety of diseases related to the brain and nervous systems， such as stroke and its sequelae， peripheral facial paralysis， rheumatoid arthritis， hypertension， and other diseases related to the motor nervous system. Its primary indications are stroke and its sequelae， followed by peripheral facial paralysis. Other new indications are gradually found. This study is expected to provide references for the clinical application of Xiao Xumingtang and the transformation of new drugs.
Abstract：Hyperthyroidism is a systemic disease characterized by clinical signs and symptoms of hypermetabolism and sympathetic nervous excitement. Based on the clinical diagnostic criteria of traditional Chinese and western medicine for hyperthyroidism，the present study summarized and evaluated animal models of hyperthyroidism. In model evaluation，the models with high coincidence degree in western medicine included the exogenous drug delivery model， the model immune to adenovirus expressing thyrotropin receptor （TSHR），the model immune to nucleic acid， and the model of yin deficiency and effulgent fire syndrome in the disease-syndrome combination. The models with high coincidence degrees in traditional Chinese medicine included the exogenous drug delivery model， the model immune to adenovirus expressing TSHR，and the model of liver-yang ascendant hyperactivity syndrome and the model of yin deficiency and effulgent fire syndrome in the disease-syndrome combination. In light of the coincidence degree， and advantages and disadvantages of traditional Chinese and western medicine，the ideal hyperthyroidism animal models are the exogenous drug delivery model， and the model immune to adenovirus expressing TSHR. In addition to the evaluation of the coincidence degree of animal models of hyperthyroidism in traditional Chinese and western medicine，this study also analyzed the advantages，disadvantages， and problems of the animal models of hyperthyroidism. Most of the animal models of hyperthyroidism were not consistent with the complexity of hyperthyroidism in clinical practice， and standardized and unified syndrome differentiation standards and four-examination information collection standards have not yet been formed. Besides， there have been few studies on the hyperthyroidism model in disease-syndrome combination in traditional Chinese medicine. To make the animal models of hyperthyroidism suitable for clinical practice，the present study proposed the improvement directions of animal models of hyperthyroidism and the necessity of promoting the evaluation system to provide a theoretical basis for the evaluation of the curative effect of Chinese medicine on hyperthyroidism， and exploration of its pharmacological action， as well as the follow-up research on the pathogenesis，prevention， and treatment of hyperthyroidism，which is expected to establish a perfect disease-syndrome model of hyperthyroidism in line with clinical characteristics of traditional Chinese and western medicine.
Keywords：hyperthyroidism;animal models;clinical disease and syndrome;traditional Chinese and Western medicine
Abstract：Hypertension is the most important risk factor for cardiovascular and cerebrovascular diseases， affecting the structures and functions of important organs of the body， such as the heart， brain， and kidney. At present， the prevalence of hypertension in China remains high. How to effectively curb the incidence of hypertension and reduce target organ damage in patients with hypertension is an urgent challenge that needs to be addressed. Traditional Chinese medicine （TCM）， by virtue of its unique efficacies and advantages， is increasingly applied around the world. In TCM， "wind" is considered as a major constant factor in the development of hypertension. Some scholars believe that hypertension is located in collaterals， and the lesions of collaterals are also important reasons for the occurrence and development of hypertension. Collateral diseases and pathogenic wind are closely related to the development of hypertension as well as target organ damage in the heart， brain， and kidney. From the six-meridian syndrome differentiation for febrile diseases， "collateral" and "wind" are closely related to the Jueyin， with collateral diseases classified into Jueyin diseases and pathogenic wind beginning in the Jueyin. The occurrence， development， and persistence of hypertension are closely related to Jueyin diseases. The present study analyzed the pathogenesis and treatment of hypertension from "collateral" and "wind" of the Jueyin， and specifically discussed the relationship between hypertension and "collateral" and "wind" of the Jueyin. It is believed that the internal depression of wind and fire in the Jueyin results in the upward impulse of liver fire， and the deficiency of Jueyin can trigger the internal movement of liver wind and stirring wind due to collateral deficiency. External contraction in the Jueyin due to the induction of external wind is the important pathogenesis of the development of hypertension and the damage to target organs such as the heart， brain， and kidney. The therapeutic methods for both "collateral" and "wind" were also discussed based on the primary prescription for Jueyin， Wumei Pills. Six-meridian syndrome differentiation can guide the therapeutic principles for all diseases and inspire posterity. Exploring the pathogenesis and treatment of hypertension from “collateral" and "wind" of the Jueyin is of great significance in guiding the prevention and treatment of hypertension， reducing target organ damage in patients with hypertension， and improving the prognosis of cardiovascular and cerebrovascular diseases.
Abstract：Traditional Chinese medicine （TCM） has played an important role in the prevention and treatment of novel coronavirus disease 2019 （COVID-19）. Qingfei Paidu decoction，as a general prescription of Diagnosis and Treatment Protocol for Novel Coronavirus Pneumonia from the sixth to eighth versions，has been proved effective clinically and is suitable for mild，moderate，severe，and critical patients. It can significantly improve clinical symptoms such as fever， cough，asthma，fatigue， etc. On the basis of the findings of relevant research papers，this paper summarized the TCM understanding of COVID-19， including etiology，pathogenesis， disease location， and treatment，and concluded that the disease is caused by the pestilential Qi，localized in the lungs， and can affect the five organs. It is mainly characterized by coldness，dampness，heat，toxicity，stasis，and deficiency. In response to the etiology and pathogenesis of the disease，the therapeutic principles at all stages are dominated by the elimination of pathogens and removal of toxicity. According to the stages of disease development，the treatment should combine the severity of the disease and the course of the disease with the TCM syndromes. Furthermore，from the clinical application of Qingfei Paidu decoction，this paper discussed the therapeutic intention of "Qingfei （clearing of lungs）" and "Paidu （removal of toxicity）". Qingfei Paidu decoction can clear the pathogenic toxin in the lungs and eliminate external pestilential Qi，which is in line with the therapeutic principles for this pandemic by regulating the triple energizer and protecting healthy Qi using both coldness and warmth to treat both the symptoms and the root cause. Additionally，the experimental research progress on Qingfei Paidu decoction and its modified prescriptions were summarized. As studied， this prescription can inhibit cytokine storm，moderate the overactive immune response，potentiate the immune function and anti-viral ability of the body，and exert its effect on COVID-19 with multiple components，multiple targets，multiple pathways， and multiple biological functions. In conclusion，Qingfei Paidu decoction，as a core prescription for the treatment of COVID-19，can rapidly contain the development of COVID-19， which has been confirmed in terms of TCM theory，clinical efficacy， and experimental research.
Keywords：novel coronavirus disease 2019 （COVID-19）;Qingfei Paidu decoction;traditional Chinese medcine theory;clinical application;experimental study
Abstract：Da Jianzhongtang， which is from Synopsis of Golden Chamber（《金匮要略》）， is one of the hundred ancient classical formulas. In this textual research， a total of 46 records on this prescription from 42 ancient books and 207 records from modern literature were screened out. Through systematic research on its name， dosage， processing， evolution of the meaning on prescription compatibility， indications， decocting methods， usage， and precautions， this paper aims to lay a literature basis for further research on Da Jianzhongtang. The results show that the converted dosages for the components are 3 g of Zanthoxyli Pericarpium， 12 g of Zingiberis Rhizoma， 6 g of Ginseng Radix et Rhizoma， and 30 g of Saccharum Granorum. The efficacy can be labeled on the prescription instruction as warming the middle and tonifying deficiency， descending adverse Qi and relieving pain. According to modern literature， the main indications include diseases in digestive system， circulatory system， orthopedic system， urinary system， and ophthalmic system， particularly the intestinal obstruction， gastric ulcer， chronic gastritis， and duodenal ulcer in the digestive system. The core pathogenesis of the above diseases is "deficiency-cold of middle energizer and internal exuberant cold". Precautions are as follows. ① For the processing of Zanthoxyli Pericarpium， stir-frying can alleviate its toxicity and increase the decoction rate of effective ingredients. ② For the dosage， overdosage will lead to dry cough， edema， etc. ③ As to the contraindications after the medication， raw， cold， or indigestible food should not be taken after medication. ④ For the syndrome taboos， this prescription should be used with caution for patents with Yin deficiency， congealing cold， and Qi stagnation， and be forbidden for patients with spleen and stomach dampness-heat. The above results lay a basis for the research and development of the compound preparation for Da Janzhongtang and provide a reference for its instructions.
Keywords：classical prescriptions;Da Janzhongtang;evolution of the meaning on prescription compatibility;efficacy;compound preparation;textual research
Abstract：Acute lung injury is a common critical respiratory disease， which can further develop into acute respiratory distress syndrome （ARDS） with a high fatality rate， but there is no effective drug at present. Phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway is involved in cell proliferation， metabolism， survival， and motility through phosphorylation of numerous downstream effector molecules. It plays an important role in the development of acute lung injury by regulating mitochondrial function， apoptosis， autophagy， oxidative stress， and inflammatory response. The active ingredients in Chinese medicinals alleviate acute lung injury by targeting the PI3K/Akt signaling pathway. There has been an explosion of research on the treatment of acute lung injury by active ingredients in Chinese medicinals via PI3K/Akt signaling pathway， which is of great clinical research value. The article presented the first summary of studies exploring the correlation between PI3K/Akt signaling pathway and acute lung injury in recent years and summed up the protective effect of the active ingredients in Chinese medicinals against acute lung injury via PI3K/Akt signaling pathway， providing innovative mindsets and strategies for clinical application of active ingredients in Chinese medicinals in the treatment of acute lung injury.
Keywords：acute lung injury;phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）;active ingredients in Chinese medicinals;autophagy;apoptosis;inflammation
Abstract：Suanzaoren Tang， as documented in the Synopsis of the Golden Chamber （《金匮要略》），consists of Ziziphi Spinosae Semen， Poria， Anemarrhenae Rhizoma， Chuanxiong Rhizoma， and Glycyrrhizae Radix et Rhizoma， and is indicated for dysphoria， consumptive disease， and insomnia. In modern clinical practice，in addition to sleep disorders，Suanzaoren Tang and its modified formulas can also be used to treat anxiety disorders， as well as insomnia，cardiac intervention，myocardial infarction， and other diseases combined with anxiety. It is also used in combination with other Chinese medicinal formulas （such as Zhizichi Tang，Ganmai Dazaotang，Baihe Zhimu Dihuangtang，and Jinlingzi San），western medicines （such as paroxetine，zopiclone，enalapril，amlodipine，metformin，and sertraline hydrochloride）， acupuncture， and acupoint application to treat anxiety， as well as cardiovascular neurosis，cancer，hypertension，type 2 diabetes， and other diseases combined with anxiety. As revealed by clinical treatment results， Suanzaoren Tang and its modified formulas can significantly reduce anxiety scores such as Hamilton Anxiety Rating Scale（HAMA） and Self-rating Anxiety Scale（SAS），and improve anxiety symptoms with significant efficacy and few side effects. As reported by experimental pharmacological studies， Suanzaoren Tang can regulate the content of neurotransmitters such as dopamine（DA），homovanillic acid（HVA），γ-aminobutyric acid（GABA），noradrenaline （NE），5-hydroxyindoleacetic acid （5-HIAA），glutamic acid（Glu），β-endorphin（β-EP），5-hydroxytryptamine（5-HT）， and nitric oxide （NO）in the brain， and mediate immune function by reducing the levels of inflammatory cytokines such as interleukin-1β（IL-1β）and tumor necrosis factor-α（TNF-α），enhancing the proliferation of B lymphocytes，phagocytosis of peritoneal macrophages， and down-regulating the proliferation of T lymphocytes. Besides， it can also regulate the endocrine level to allow the homeostasis of nervous system， endocrine system， and immune system by increasing adrenocorticotropic hormone（ACTH）and corticosterone（CORT）levels and up-regulating glucocorticoid receptor（GR）expression， finally achieving the effect of anxiety prevention and treatment. The present study systematically reviewed the clinical and basic research progress on the prevention and treatment of anxiety with Suanzaoren Tang to provide references for the research on the mechanism and material basis of Suanzaoren Tang and the development of new drugs for anxiety.
Abstract：Follicular helper T （Tfh） cells are a newly discovered subset of CD4+ T cells. As reported， abnormalities in their development， differentiation， and function are closely related to the occurrence of autoimmune diseases. Psoriasis is an autoimmune skin disease and it is intractable with a prolonged course. At present， it is generally believed that immune imbalance mediated by T cells is the core mechanism of the pathogenesis of psoriasis. In the context of this mechanism， Tfh cells are associated with psoriasis， and their cellular level and abnormal expression of related candidates can promote the occurrence of psoriasis. In terms of treatment， Chinese medicine， by virtue of the characteristics of wide application and low price， serves as a good complementary and alternative treatment option for psoriasis. As confirmed by previous findings， some active ingredients or preparations of Chinese medicine used in the treatment of psoriasis can also intervene in and regulate the immune response mediated by Tfh cells and the related candidates. Based on the research reports and experimental data， the present study reviewed the research progress from the differentiation of Tfh cells， the relationship between Tfh cells and psoriasis， and the intervention and regulation of Tfh cells and related molecules by Chinese medicine， which is expected to provide certain theoretical support and references for the determination of new strategies for psoriasis treatment and research in related fields.
Keywords：follicular helper T cells;psoriasis;traditional Chinese medicine;molecules;review
Abstract：Dysmenorrhea is a common gynecological disease in clinic， with primary dysmenorrhea and secondary dysmenorrhea. Primary dysmenorrhea is usually not accompanied by organic lesions in genital organs， which is mainly related to the increase of prostaglandin content in endometrium during menstruation. Secondary dysmenorrhea is accompanied by organic lesions of reproductive organs， often associated with local lesions of reproductive organs， but also with patients' mental factors and neuroendocrine factors.The incidence of dysmenorrhea is as high as 73.8%， and there is no radical cure method， which has a great impact on the life， work and learning of patients. Chinese medicine essential oil widely exists in aromatic Chinese medicine， with antibacterial， antioxidant and anticancer activities. It can regulate neuroendocrine function， anti-inflammatory and analgesic effects， and improve mood by regulating the levels of prostaglandins， oxytocin and other hormones in the body and regulating the emotions of patients， there by alleviating dysmenorrhea to a certain extent. In recent years， many scholars have made more in-depth research on Chinese medicine essential oil in alleviating dysmenorrhea， but there is a lack of comprehensive collation of such studies. In this regard， the author has systematically sorted out the generation and classification of dysmenorrhea， the mechanism of action of essential oil of traditional Chinese medicine to alleviate dysmenorrhea and the application of essential oil of traditional Chinese medicine in the field of dysmenorrhea by consulting relevant literature in Chinese and foreign languages in recent years， so as to provide reference for the treatment of dysmenorrhea.
Keywords：essential oils of traditional Chinese medicine;primary dysmenorrhea;secondary dysmenorrhea;action mechanism;application
Abstract：Sophorae Tonkinensis Radix et Rhizoma （STRER） is a commonly used Chinese medicine in clinical practice， which has the effects of clearing heat， removing the toxin， alleviating edema， and relieving sore throat. In recent years， the clinical reports of STRER-induced poisoning have gradually increased， with neurotoxicity and hepatotoxicity as the main characteristics of the acute attack. Timely treatment will lead to the good prognosis， but long-term or high-dose administration will cause irreversible damage. Therefore， the safety of clinical use of STRER should be highlighted. The chemical components in STRER mainly include alkaloids， flavonoids， triterpenoids， triterpenoid saponins， and polysaccharides， as well as small amounts of proteins， organic acids， and trace elements， where alkaloids both serve as the important material basis for the pharmacodynamic action and the main substances causing toxicity. The adverse events induced by STRER and its alkaloids include nerve injury, Hepatic injury, cardiovascular injury, kidney injury and reproductive injury， and gastrointestinal reaction. Quinolizidine alkaloids are the main toxic components， mainly including matrine， oxymatrine， cytisine， sophocarpine， oxysophocarpine， sophoridine， sophoramine， and lehmannine. Many studies have been carried out on the toxicity of different extracts and alkaloids of STRER in China and abroad， but there are no comprehensive and detailed reports on the toxicity mechanism of alkaloids in STRER. As a Chinese medicine， STRER is widely used. It's an urgent problem to clarify the material basis and mechanism of toxicity caused by STRER and reduce the toxicity for good clinical application. The present study reviewed the components of alkaloids， toxicity， and toxic mechanism of extracts and alkaloids in STRER to provide the basis for further development and clinical safe and effective application of STRER.
Keywords：Sophorae Tonkinensis Radix et Rhizoma;alkaloids;component;adverse reaction;mechanism
Abstract：Andrographolide， a diterpene lactone， is the important material basis for the pharmacological effect of the Chinese medicinal Andrographis paniculata （Burm.f.）Nees. Modern pharmacological research has shown that andrographolide has many pharmacological activities such as anti-inflammation， bacteriostat， anti-virus， anti-tumor， protecting liver， promoting the function of gallbladder， and protecting the cardiovascular system and nervous system. It has significant anti-inflammatory activity which involves multiple targets. To be specific， it can inhibit nuclear factor-κB （NF-κB）， signal transduction and activator of transcription 3 （STAT3）， and other signaling pathways， reduce the synthesis and release of downstream inflammatory mediators， and regulate oxidative stress and immune response to achieve anti-inflammatory effect on various inflammatory diseases. At the same time， it suppresses a variety of tumor cells by inhibiting tumor cell proliferation， blocking cell cycle， and inducing tumor cell apoptosis. Its anti-tumor mechanism involves cellular signaling pathways such as Notch， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， NF-κB， and secreted glycoprotein/β-catenin （Wnt/β-catenin）. In addition， it can also alleviate diabetes by regulating glucose metabolism. According to related research， it often exerts pharmacological effects through multiple pathways and multiple targets， but the specific targets are unclear. Therefore， this article summarizes the relevant studies on the pharmacological effects and mechanisms of andrographolide in the past three years and puts forward the future research directions， which is expected to serve as a reference for the further in-depth research and development and utilization of andrographolide.