Abstract:ObjectiveTo investigate the effects of Daizongfang (DZF) on insulin resistance (IR) of adipocytes induced by different methods.MethodThe cocktail induction method was adopted to induce the differentiation and maturity of 3T3-L1 preadipocytes. An IR model in mature adipocytes was established by the induction of palmitic acid (PA), high-concentration glucose (HG), and dexamethasone (DEX). DZF extracts at different concentrations (2.0, 0.5, 0.1 g·L-1) intervened for 24 hours. A model group, a rosiglitazone (RSG) group, and a blank control group were set up at the same time. The glucose concentration in the culture supernatant was measured by the glucose oxidase-peroxidase (GOD-POD) method. Glucose consumptions under basic conditions (GBasic) and insulin stimulation (GIns) were calculated to evaluate the insulin sensitivity index (ISI). The mRNA expression of glucose transporter 4 (GLUT4) was detected by the real-time polymerase chain reaction (PCR).ResultCompared with the model group, the DZF (2.0 g·L-1) showed increased GBasic, GIns, and ISI in three IR models (P<0.05, P<0.01). In addition, for the PA-induced IR model, GBasic and GIns in the DZF (0.5 g·L-1) group were elevated (P<0.01), and GBasic, GIns, and ISI in the RSG group increased (P<0.05, P<0.01). For the HG-induced IR model, GIns and ISI increased in the DZF (0.5 g·L-1) group (P<0.05), and GBasic, GIns, and ISI were elevated in the RSG group (P<0.01). For the DEX-induced IR model, GIns and ISI increased in the RSG group (P<0.01). In the three models, there were differences among groups with different doses. GBasic, GIns, and ISI in the high-dose DZF group increased in varying degrees compared with those in the medium- and low-dose DZF groups (P<0.05). In the three models, the DZF (2.0 g·L-1) group and the RSG group both increased GLUT4 mRNA expression (P<0.05).ConclusionDZF can reduce IR of adipocytes induced by HG, DEX, or PA in a dose-dependent manner and increase glucose uptake in an insulin-independent manner, which may be related to the increase in GLUT4 expression.
Abstract:ObjectiveTo establish high performance liquid chromatography (HPLC) fingerprint and multi-component determination for the substance benchmark of Yiweitang, and to evaluate its quality in combination with chemical pattern recognition method.MethodFifteen batches of substance benchmark of Yiweitang were prepared, the "Chinese medicine chromatographic fingerprint similarity evaluation system" (2012 edition) was used to calculate similarity. Cluster analysis, principal component analysis and orthogonal partial least squares-discriminant analysis were employed to handle the common peaks for evaluating the quality difference among 15 batches of the substance benchmark. The contents of catalpol, verbascoside and methylophiopogonanone A were determined with mobile phase system of acetonitrile-phosphoric acid solution at detection wavelengths of 210 nm and 334 nm.ResultThere were 22 common peaks in HPLC fingerprint of the substance benchmark, among them, peaks 1, 9, 12, 14-17, 19 and 20 belonged to Rehmanniae Radix, peaks 3, 4, 6, 7 and 21 belonged to Glehniae Radix, peaks 5 and 22 belonged to Ophiopogonis Radix, peaks 2 and 18 belonged to Polygonati Odorati Rhiaoma, peak 8 was the common peak of Ophiopogonis Radix and Rehmanniae Radix, peak 10 was shared by Ophiopogonis Radix, Polygonati Odorati Rhiaoma and Rehmanniae Radix, peak 11 was the common peak of these four herbs, and peak 13 was shared by Polygonati Odorati Rhiaoma and Rehmanniae Radix. The similarities between HPLC fingerprints of 15 batches of the substance benchmark and the control fingerprint were all >0.90, the samples could be divided into four categories by three chemical pattern recognition methods. Quantitative analysis showed that the contents of catalpol, verbascoside and methylophiopogonanone A among 15 batches of samples ranged from 0.37% to 1.14%, 0.002% to 0.054% and 0.016% to 0.079%, respectively.ConclusionThe established fingerprint and determination for the substance benchmark of Yiweitang have good separation and high accuracy, which reflect the overall chemical composition characteristics of Yiweitang, and can provide experimental basis for the further development and quality control of the compound preparations of this famous classical formula.
Abstract:ObjectiveTo observe the effect of Fangji Huangqitang (FJHQT) on collagen induced arthritis (CIA) and synovial angiogenesis in DBA/1 mice.MethodDBA/1 mice were randomly divided into normal group, CIA group and FJHQT group. DBA/1 mice in CIA group and FJHQT group were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day, and DBA/1 mice were immunized with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day to establish CIA model. On the day of the second immunization, the drug was given by gavage once a day for 28 days. On the 22nd day, the arthritis score and other symptoms of CIA mice were observed. On the 49th day, Hematoxylin eosin (HE) staining was carried out to observe the angiogenesis in the synovium of CIA mice, the expression of vascular endothelial cell marker platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial growth factor (VEGF) in the synovium of CIA mice were detected. Immunofluorescence double staining was used to detect the mature and immature vessels in the synovium of CIA mice. And the microvascular growth of the rat thoracic aortic ring was induced by VEGF (20 μg·L-1). The effects of FJHQT (0.25, 0.5, 1 g·L-1) at different concentrations were observed under microscope.ResultCompared with the normal group, the inflammation, joints, red and swelling of the inflammatory joints of the CIA group were significantly increased (P<0.01). The scores of clinical arthritis, the incidence rate, synovial inflammation and angiogenesis were significantly increased (P<0.01). The density of blood vessels, the positive expression of CD31 and VEGF, the number of immature vessels in synovial membrane were significantly increased (P<0.01). And compared with the CIA group, the inflammation, joint swelling, and malformation of the FJHQT group were significantly improved, the clinical arthritis score, incidence rate, synovial inflammation and angiogenesis were significantly reduced (P<0.01). The vascular density, the positive expression of CD31 and VEGF, and the number of immature blood vessels in synovial membrane were significantly increased (P<0.01). Compared with blank group, VEGF could significantly induce the growth of microvasculature in rat thoracic aortic ring (P<0.01). Compared with VEGF group, FJHQT(0.25, 0.5, 1 g·L-1) could significantly inhibit the formation of microvasculature in rat thoracic aortic ring (P<0.01).ConclusionFJHQT can effectively alleviate the clinical symptoms and condition of CIA mice, reduce the clinical arthritis score and incidence rate,and inhibit the synovial angiogenesis of CIA mice joints and VEGF induced microvascular formation in rat thoracic aortic rings.
Abstract:ObjectiveTo explore the efficacy and mechanism of Danggui Shaoyaosan on rats of nonalcoholic fatty liver disease (NAFLD).MethodSixrty SPF SD male rats were randomly divided into normal group, model group,essentiale (0.144 g·kg-1) and low, middle and high-dose of Danggui Shaoyaosan groups (2.44, 4.88, 9.76 g·kg-1). High fat diet were fed to bulid the NAFLD model, and each treatment group was given corresponding drugs at the same time. After 8 weeks, the serum and liver tissue were collected to detect the contents or activities of total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartic acid aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in serum, the contents of TC, TG and free fatty acid (FFA) in liver tissue, Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to observe the gene and protein expressions of Toll-like receptor 4 (TLR4), myeloid different factory 88 (MyD88) and c-Jun n-terminal kinase (JNK) and the protein expression of phosphorylation JNK(p-JNK) in liver tissue. Hematoxylin-eosin (HE) staining and Oil red staining to observe the pathological morphological changes of liver.ResultCompared with control group, the contents or activities of TC, TG, ALT, AST and TNF-α in serum, the contents of TC, TG and FFA in liver and the gene and protein expressions of TLR4, MyD88, JNK, and the protein expression of p-JNK in liver tissue of model group were distinctly increased (P<0.01), the content of IL-10 was significantly decreased (P<0.01). Compared with model group, the contents or activities of TC, TG, ALT, AST and TNF-α in serum, the contents of TC, TG and FFA in liver and the mRNA and protein expressions of TLR4, MyD88 and JNK, and the protein expression of p-JNK in liver tissue of Danggui Shaoyaosan groups were significantly reduced (P<0.05,P<0.01), the content of IL-10 in serum of Danggui Shaoyaosan groups was distinctly increased(P<0.05,P<0.01), HE staining and Oil red staining show that the degree of liver steatosis was alleviated obviously by Danggui Shaoyaosan.ConclusionDanggui Shaoyaosan has a better treatment on NAFLD by inhibiting TLR4/MyD88/JNK pathway and alleviating the inflammation response.
Abstract:ObjectiveTo observe the effects of Albiziae Flos (AF) and Polygalae Radix (PR) alone and their combination on the improvement of depression-like behavior in rats with chronic unpredictable stress (CUS) as well as on hippocampal ultrastructure and the expression of cyclic adenosine monophosphate response element binding protein (CREB) and nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2), to explore their action mechanisms.MethodSeventy-two Wistar rats were randomly divided into the normal group, model group, AF group, PR group, AF-PR group, and fluoxetine group. Rats in all groups except for the normal group were exposed to CUS and separated feeding to induce depression. Since the first day of modeling, rats in the AF group, PR group, AF-PR group were provided with the corresponding decoction containing 1.05 g·kg-1 total crude drug by gavage, the ones in the fluoxetine group with 2.1 mg·kg-1 fluoxetine hydrochloride aqueous solution, and those in the normal group and model group with the distilled water, for 28 successive days. The open field test and forced swimming test were performed 1 d before modeling and 7, 14, 21, 28 d after modeling, respectively. The morphological changes in hippocampus were observed under an electron microscope on the 28th day. The superoxide dismutase (SOD) and malondialdehyde (MDA) levels in hippocampus were detected by ultraviolet spectrophotometry, and the expression levels of CREB and NOX2 were detected by real-time polymerase chain reaction (Real-time PCR) and Western blot.ResultThe behavioral experiment results showed that the number of horizontal activities and sugar water consumption in the model group declined as compared with those in the normal group, while the immobility time in the forced swimming test was prolonged (P<0.05, P<0.01). Compared with the model group, the AF group, PR group, and AF-PR group exhibited elevated number of horizontal activities, increased sugar water consumption but shortened immobility time in the forced swimming test (P<0.05, P<0.01). Compared with the AF group or PR group, the AF-PR group showed significantly different behavioral indexes (P<0.05). Morphological results showed that the mitochondria of the model group were obviously swollen and the ultrastructure of the hippocampus was destroyed. By contrast, the hippocampal ultrastructure in each administration group was close to normal. The comparison with the normal group revealed that the activity of SOD in the hippocampus of the model group was significantly reduced, whereas the content of MDA was elevated (P<0.01). Compared with the model group, the AF group, PR group, and AF-PR group displayed increased activity of SOD and decreased content of MDA in the hippocampal tissues (P<0.05, P<0.01). Compared with AF or PR alone, the herbal pair AF-PR resulted in significant differences in the above-mentioned indexes (P<0.05, P<0.01). The results of Real-time PCR and Western blot demonstrated that NOX2 expression in the hippocampus of the model group was up-regulated in comparison with that in the normal group, while the CREB expression was down-regulated (P<0.05, P<0.01). Compared with the model group, the AF group, PR group, and AF-PR group all showed diminished NOX2 expression but elevated CREB expression in the hippocampal tissues (P<0.05, P<0.01). The protein expression levels of NOX2 and CREB in the AF group or PR group were significantly different from those in the AF-PR group (P<0.01).ConclusionAF and PR alone and their combination improve the depression-like behavior of rats exposed to CUS, which may be related to the reduction of oxidative stress, the up-regulation of CREB expression, and the down-regulation of NOX2 expression in hippocampus.
Keywords:Albiziae Flos;Polygalae Radix;herbal pair;depression;behavioral experiment;cyclic adenosine monophosphate response element binding protein (CREB);nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2)
Abstract:ObjectiveTo investigate the therapeutic effect of polydatin on ulcerative colitis (UC) in mice and its regulation of protein kinase Cθ(PKCθ)/signal transducer and activator of transcription 3(STAT3) signaling on T helper cell 17(Th17) and its mechanism in the treatment of UC.MethodThe 32 male C57BL/6 mice were randomly divided into normal group, model group, polydatin group (0.045 g·kg-1) and sulfasalazine group (0.5 g·kg-1). The UC model was established by giving 3% dextran sodium sulfate (DSS) solution to free drinking water in mice. Polydatin and sulfasalazine groups were given by gavage 0.5 h before modeling for 7 days. The normal group and model group were given the same amount of normal saline. After the last administration, the colonic tissue was taken and hematoxylin-eosin (HE) was used to observe the pathological changes of colonic tissue. Flow cytometry was used to detect the proportion of Th17 in the lamina propria of colonic mucosa. The expression of interleukin-17A (IL-17A) in serum was detected by enzyme-linked immunosorbent assay (ELISA). Polydatin was added to CD4+ T cells purified from spleen of C57BL/6 mice by magnetic-activated cell sorting (MACS) under the stimulation of cell stimulation cocktail in vitro in order to detect its impact on PKCθ and STAT3 phosphorylation.ResultCompared with normal group, the body weight was significantly decreased, and disease activity index (DAI) scores of the model group was significantly increased (P<0.01), the colonic mucosal epithelium was damaged and inflammatory cells infiltration in the mucosa and submucosa was obvious, the proportion of Th17 in the lamina propria of colonic mucosa was significantly increased (P<0.01), and the content of serum IL-17A was significantly increased (P<0.01). Compared with the model group, the weight and DAI score of polydatin and sulfasalazine groups were significantly improved (P<0.01), the degree of colon tissue damage was significantly improved, the proportion of Th17 in colon mucosa lamina propria was significantly decreased (P<0.01), and the content of IL-17A in serum was significantly decreased (P<0.01). In vitro experiments showed that polydatin could significantly inhibit the phosphorylation of PKCθ and STAT3 in Th17 (P<0.01) as well as IL-17A secretion.ConclusionPolydatin can improve the ulcerative colitis in mice via inhibiting the phosphorylation of PKCθ and STAT3 to preclude IL-17A secreting in Th17.
Keywords:polydatin;ulcerative colitis;protein kinase Cθ(PKCθ);signal transducer and activator of transcription 3(STAT3);T helper cell 17(Th17);interleukin-17A(IL-17A)
Abstract:ObjectiveTo compare the characteristics of four commonly adopted animal models of hyperuricemia (HUA) for traditional Chinese medicine (TCM) screening, so as to choose the adequate model for screening Chinese herbs and herbal compounds capable of lowering the uric acid.MethodFifty-four male SD rats were randomly divided into nine groups, namely the normal group, hypoxanthine (HX) + oxonic acid potassium salt (OAPS) model group, yeast extract (YE) + OAPS model group, low-dose adenine (AD) + ethambutol (EMB) model group, high-dose AD + EMB model group, and four positive drug allopurinol (Allo) groups. The modeling lasted for 14 d. The levels of serum uric acid (SUA), urinary uric acid (UUA), serum creatinine (SCr), urea nitrogen (BUN), kidney injury molecule 1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) were detected on the 3rd, 7th, and 14th days. Urine was collected on the 7th and 14th days to investigate changes in urine volume, and the crystals in the residual urine were observed under a polarizing microscope. After the modeling, the kidney was harvested and weighed, followed by pathological examination.ResultThe urine volumes in the HX + OAPS model group and high-dose AD + EMB model group were significantly reduced (P<0.05). The renal indexes of each model group, except for the YE + OAPS model group, were significantly elevated (P<0.05, P<0.01). The increase in SUA of the HX + OAPS model group and YE + OAPS model group started later (P<0.05). The KIM-1 and NGAL levels of the HX + OAPS model group rose significantly from the 7th day (P<0.05, P<0.01), and the BUN increased significantly on the 14th day (P<0.05). There was no significant difference in the above-mentioned indicators in the YE + OAPS model group. The SUA levels of the low- and high-dose AD + EMB model groups increased significantly on the 3rd day (P<0.05, P<0.01), with a persistent increase found in the low-dose AD + EMB model group. Besides, the increase in BUN, KIM-1, and NGAL occurred later (P<0.05, P<0.01). By contrast, the high-dose AD + EMB model group exhibited a transient increase in SUA. Moreover, the SCr, BUN, KIM-1, and NGAL elevation occurred earlier and were more obvious than those in the low-dose AD + EMB model group (P<0.01). Remarkable histomorphological abnormalities were detected in the kidney of all model groups, except for the YE+OAPS model group, with the most severe injury present in the high-dose AD+EMB model group.ConclusionThe four models commonly used to screen TCM have their own characteristics. In the four models, the SUA elevation in the HX + OAPS model group and YE + OAPS model group started later, with the mild renal injury observed in the HX + OAPS model group instead of the YE + OAPS model group. The SUA of the low-dose AD + EMB model group increased rapidly and lasted for a long time, accompanied by mild renal injury. The SUA of the high-dose AD + EMB model group only showed a transient increase, accompanied by severe renal injury. The investigation on the characteristics and application of different models and the evaluation of these models based on sensitive and objective indicators are helpful for determining the suitable model for the screening of TCM targeting HUA in the future.
Keywords:hyperuricemia (HUA);traditional Chinese medicine (TCM);animal model;renal injury;urate crystals
Abstract:ObjectiveTo observe the effects of Huazhuo Jiedu Shugan Prescription (HZJDSG) on learning, memory, and the expression of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway-related proteins in epileptic rats, and to explore its possible mechanism.MethodForty-eight SPF male SD rats were randomly divided into a normal group, a model group, a sodium valproate (0.19 g·kg-1) group, and low- (2.7 g·kg-1), medium- (5.4 g·kg-1), and high-dose (10.8 g·kg-1) HZJDSG groups, with eight rats in each group. The normal group received 0.9% sodium chloride solution (0.035 g·kg-1) by intraperitoneal injection, and the other five groups received pentetrazol (PTZ) at the same dose to induce a chronic epilepsy model for a total of 14 times. The drug groups received corresponding drugs and the normal group and the model group received 0.9% sodium chloride solution at the same volume once a day for 28 days. During the drug intervention period, epilepsy was maintained in each modeling group by intraperitoneal injection of PTZ on day 7, 14, 21, and 28. The behavioral changes of rats were observed by Morris water maze and the pathomorphological changes of rat hippocampal neurons by hematoxylin-eosin (HE) staining. The protein expression of phosphorylation Akt(p-Akt)and p-GSK-3β was detected by immunohistochemistry and the protein expression of PI3K, Akt, p-Akt, GSK-3β, and p-GSK-3β by Western blot.ResultCompared with the normal group, the model group showed prolonged platform finding time (P<0.01), reduced number of platform crossings (P<0.01), structural damage of neurons in the CA1 region of the hippocampus, down-regulated protein expression of p-Akt and p-GSK-3β in the CA1 region of the hippocampus (P<0.05), and reduced relative expression of PI3K, p-Akt, and p-GSK-3β in the hippocampus (P<0.01). Compared with the model group, the sodium valproate group and the HZJDSG groups showed shortened platform finding time (P<0.01) and improved neuronal structure in the CA1 region of the hippocampus, while the sodium valproate group and the high- and medium-dose HZJDSG groups exhibited increased number of platform crossings (P<0.01), up-regulated protein expression of p-Akt and p-GSK-3β in the CA1 region of the hippocampus (P<0.05), and elevated relative expression of PI3K, p-Akt, and p-GSK-3β (P<0.01).ConclusionHZJDSG can improve the learning and memory of epileptic rats, and its antiepileptic effect may be achieved by the activation of PI3K/Akt/GSK-3β pathway-related proteins.
Keywords:Huazhuo Jiedu Shugan prescription;epilepsy;rat;learning and memory;phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) signaling pathway
Abstract:ObjectiveTo study the effect of Wenjing Huayu Zhitong therapy (Xiangyan Zhitong prescription, XZP) on the mitogen activated protein kinase (MAPK)/extracelluar regulated protein kinase (ERK) signaling pathway of primary dysmenorrhea (PD) rats, and explore the pathogenesis of PD and the mechanism of action of Wenjing Huayu Zhitong therapy.MethodForty-eight female SPF-grade Wistar rats were randomly divided into blank group,model group, western medicine group, low-dose XZP group, medium-dose XZP group, and high-dose XZP group, with 8 rats in each group. In addition to the blank group, dysmenorrhea rat model with cold coagulation and blood stasis syndrome was established by cold stimulation combined with estradiol benzoate and oxytocin. The rats in the blank group,model group,western medicine group, low-dose XZP group, medium-dose XZP group, and high-dose XZP group were given distilled water, distilled water,0.06 g·kg-1 ibuprofen, 6.55 g·kg-1 XZP, 13.09 g·kg-1 XZP, and 26.18 g·kg-1 XZP, respectively, by gavage for 6 days. The writhing latency and writhing frequency of rats were recorded within 30 min after oxytocin injection.Western blot was used to detect the protein expression of B-Raf, mitogen activates extracellular regulated kinases1/2 (MEK1/2), extracellular regulated kinases1/2 (ERK1/2), p-MEK1/2, p-ERK1/2, c-Jun, and cyclooxygenase-2 (COX-2) in rat uterus. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to detect the mRNA expression of B-Raf, MEK1, MEK2, ERK1, ERK2, c-Jun, and cyclooxygenase-2 (COX-2) in rat uterus.ResultCompared with the model group,the treatment groups showed insignificantly prolonged writhing latency and significantly reduced writhing frequency (P<0.01). On the 6th day of modeling, there was no significant difference in the quantitative scores of symptoms and signs among the treatment groups. On the 12th day of modeling, the scores changed little in the western medicine group and the low-dose XZP group and decreased significantly in the medium- and high-dose XZP groups (P<0.01) compared with those in the model group. Compared with those in the blank group, the protein and mRNA levels of p-MEK1/2, p-ERK, B-Raf, c-Jun, and COX-2 in the model group were significantly up-regulated (P<0.01). Compared with those in the model group, the protein and mRNA levels of p-MEK1/2, p-ERK1/2, B-Raf, c-Jun, and COX-2 in the western medicine group, medium-dose XZP group, and high-dose XZP group were significantly down-regulated (P<0.05,P<0.01).ConclusionThe mechanism of Wenjing Huayu Zhitong therapy in treating PD with cold coagulation and blood stasis syndrome may be related to the down-regulation of MAPK/ERK signaling pathway.
Keywords:primary dysmenorrhea;cold coagulation and blood stasis syndrome;Wenjing Huayu Zhitong therapy;mitogen activated protein kinase (MAPK)/extracelluar regulated protein kinase (ERK) signaling pathway
Abstract:ObjectiveTo investigate the effects of capsaicin on colon cancer SW480 cells and the underlying molecular mechanism through the transient receptor potential vanilloid 1(TRPV1).MethodCapsaicin groups with different concentrations and a blank group were set up. The cell viability was detected by cell counting kit-8 (CCK-8) after SW480 cells were treated with capsaicin(50,100,200,300,400,500,600,800,1 000 μmol·L-1) for 12,24,and 48 h to select the concentration of capsaicin which can effectively inhibit proliferation. The cell cycle and apoptosis were detected by flow cytometry after SW480 cells were treated with capsaicin (200,400,800 μmol·L-1) for 24 h. The protein expression levels of TRPV1,p53,p-p53,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved cysteinyl aspartate-specific protease-3(cleaved Caspase-3),cleaved Caspase-8,and cleaved poly adenosine diphosphate ribose polymerase (PARP) were detected by Western blot after SW480 cells were treated with capsaicin (200,400 μmol·L-1) for 24 h.In addition,the apoptosis was detected after SW480 cells were treated with TRPV1 microRNA(mRNA) and capsaicin(200 μmol·L-1). Western blot analysis was used to detect the protein expression levels of the above proteins.ResultAs compared with the blank group,capsaicin(≥200 μmol·L-1)significantly inhibited the cell viability of SW480 cells(P<0.01) in dose- and time-dependent manners. The cell cycle was arrested in G2/M phase by 200 and 400 μmol·L-1 capsaicin treatment,and arrested in G1 phase by 800 μmol·L-1 capsaicin treatment (P<0.05). Flow cytometry showed that capsaicin (200, 400, 800 μmol·L-1) significantly promoted apoptosis of SW480 cells simultaneously(P<0.05,P<0.01). Western blot showed that capsaicin (200,400 μmol·L-1) significantly up-regulated the protein levels of apoptosis-related proteins(p53,p-p53,Bax,cleaved Caspase-3,cleaved Caspase-8,and cleaved PARP) (P<0.05,P<0.01),and significantly down-regulated Bcl-2(P<0.01). In addition,siRNA-mediated knockdown of TRPV1 significantly attenuated capsaicin-induced apoptosis and the protein levels of apoptosis-related proteins in SW480 cells(P<0.05,P<0.01).ConclusionCapsaicin can inhibit cell proliferation,arrest cell cycle,and induce apoptosis of SW480 cells,and the possible mechanism may be related to TRPV1 activation.
Abstract:ObjectiveTo observe the effects of artesunate (ART) on experimental choroidal neovascularization (CNV) and the expression of related proteins, and to explore the underlying mechanism.MethodEighty BN rats were randomly divided into five groups: a normal group, a model group, a conbercept group, and low- (10.08 mg·kg-1·d-1) and high-dose (20.16 mg·kg-1·d-1) ART groups, with 16 rats in each group. A CNV model was established with 532 nm laser in rats of the groups except for the normal group. The rats in each group were treated with corresponding drugs by gavage for 14 days. The normal group, the model group, and the conbercept group received 1% CMC-Na solution at the same volume, while the conbercept group received an intravitreal injection (5 μL) once. On days 7 and 14, fundus fluorescein angiography (FFA) was used to evaluate the fluorescein leakage (gray value) of CNV. Hematoxylin-eosin (HE) staining was adopted to observe the histopathological changes of CNV. Western blot was employed to detect the protein expression levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in the retina and choroid.ResultFFA results showed that compared with the normal group, other groups showed increased gray value on days 7 and 14 (P<0.01). On day 7, the gray value of the high-dose ART group and the conbercept group decreased compared with that in the model group (P<0.01). On day 14, the gray value of the ART groups and the conbercept group decreased in varying degrees compared with that in the model group (P<0.05, P<0.01). HE results showed that compared with the normal group, the model group showed increased thickness of CNV on days 7 and 14 (P<0.01). Compared with the model group, the ART groups and the conbercept group displayed decreased thickness of CNV (P<0.01). Western blot results revealed that the expression of HIF-1α and VEGF in the model group increased in varying degrees on the days 7 and 14 compared with that in the normal group (P<0.05, P<0.01), while compared with the model group, the ART groups and the conbercept group showed decreased expression (P<0.01).ConclusionART can inhibit experimental CNV by down-regulating the expression of HIF-1α and VEGF in the early stage of experimental CNV formation.
Abstract:ObjectiveTo investigate the possible mechanism of Xieheyin in alleviating obese polycystic ovary syndrome with insulin resistance(PCOS-IR)and reducing inflammatory response.MethodTen of sixty SPF femlae C57BL/6J mice were randomly selected as the normal group,and the rest mice were given letrozole 0.002 g·kg-1 combined with fecal suspension 2 g·kg-1 for 28 consecutive days to establish model of PCOS-IR.The mice that were successfully modeled were randomized into the model group,metformin group(0.25 g·kg-1),and low(10 g·kg-1),medium(20 g·kg-1),and high-dose(40 g·kg-1)Xieheyin groups,and administered with the corresponding drugs by gavage,once a day,for four consecutive weeks. Except the normal control group, the mice in the other groups were continuously given fecal suspension combined with letrozole solution to maintain the model during the treatment. The mice were weighed once a week.Levels of fasting blood glucose (FBG) were detected by blood glucose test strips.And enzyme-linked immunosorbent assay (ELISA) method was used to detect serum testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),fasting insulin(FINS)level,and LH/FSH and Homeostasis model assesment of insulin resistance (HOMA-IR) were calculated.The uterus and ovaries were weighed and fixed.Hematoxylin-eosin(HE)staining was used to observe ovarian tissue pathology morphology. Western blot was used to detect the expression levels of tight junction key molecular zonula occludens 1(ZO-1),occludin in colon tissues,and the expression levels of Toll-like receptor 4/nuclear factor kappa B/Nod-like receptor protein 3(TLR4/NF-κB/NLRP3)signaling pathway and inflammation associated proteins cysteinyl aspartate specific proteinase-1(Caspase-1) and interleukin-1β(IL-1β) in colon tissues.ResultCompared with normal control group,the body weight of mice in the model control group increased significantly(P<0.01). Serum FINS,FBG,HOMA-IR,T,LH/FSH were significantly increased(P<0.05,P<0.01). The uterine organ ratio were decreased significantly(P<0.01),while the ovarian organ ratio were significantly increased(P<0.01). The number of atresia follicles and cystic dilatation follicles increased significantly,and the number of corpus luteum significantly decreased,the thickness of follicular granulosa cells also decreased,while the white membrane thickness of the ovary increased. Tight junction related ZO-1,occludin proteins in colon tissues were all decreased(P<0.01).The relative expression levels of inflammation-related protein IL-1β,Caspase-1 and TLR4/NF-κB/NLRP3 target protein signaling pathway were significantly increased(P<0.05).Compared with model control group, the body weight of mice in the low,middle and high dose Xieheyin group decreased significantly(P<0.01). The serum T,LH/FSH,FINS,FBG,HOMA-IR were significantly decreased(P<0.05,P<0.01). The uterine organ ratio were increased(P<0.05),while the ovarian organ ratio were decreased(P<0.05). The number of cystic follicles decreased and corpus luteum increased,the thickness of follicular granulosa cells increased and be arranged normally,while the white membrane thickness of the ovary increased slightly. The expressions of ZO-1,occludin proteins were increased(P<0.01). The expression levels of IL-1β,Caspase-1 and TLR4/NF-κB/NLRP3 target protein in the high dose group were significantly decreased(P<0.01).ConclusionXieheyin could activate intestinal TLR4/NF-κB/NLRP3 signaling pathway,inhibit pro-inflammatory factor secretion,improve obesity and IR,which was correlated with rebuilding intestinal mucosal barrier and inhibiting intestinal inflammation.
Abstract:ObjectiveTo explore the clinical efficacy of modified Zhizhutang in treating senile primary constipation due to Qi-Yin deficiency and its influences on gastrointestinal neurotransmitters and gastrointestinal motility.MethodOne hundred and twenty-one eligible patients were randomly divided into a control group (n=60) and an observation group (n=61). On the basis of lifestyle adjustments, patients in the control group further received oral Qihuang Tongmi soft capsule,three capsules per time,three times per day, whereas those in the observation group were further treated with modified Zhizhutang,one bag per day, for eight successive weeks. The number of complete spontaneous bowel movements (CSBM, normal defecation was considered if CSBM ≥ three times per week) before and after treatment was recorded. The main symptoms of constipation,the patient assessment of constipation quality of life (PAC-QOL), and the traditional Chinese medicine (TCM) syndrome before and after treatment were scored, followed by the anorectal pressure measurement and colonic transit test before and after treatment. The levels of somatostatin (SS),motilin (MTL),substance P (SP) and vasoactive intestinal peptide (VIP) before and after treatment were detected.ResultThe weekly average number of CSBM in the observation group was greater than that of the control group (P<0.01), but the TCM syndrome score was lower than that of the control group (P<0.01). The normal defecation rate in the observation group was 57.38% (35/61),which was higher than 35% (21/60) of the control group (χ2=6.092,P<0.05). The scores of the main symptoms of constipation in the observation group were significantly lower than those in the control group (P<0.01). The PAC-QOL subscale scores and the total score of the observation group significantly declined as compared with those of the control group (P<0.01). The anal resting pressure and anal maximum systolic pressure of the observation group were higher than those of the control group (P<0.01),while the first sensation volume, urge sensation volume and maximum tolerable volume were lower (P<0.01). The excretion rates of markers in the observation group at 24 h,48 h and 72 h were significantly elevated in comparison with those in the control group (P<0.01). Compared with the control group, the observation group also exhibited increased serum MTL and SP (P<0.01) and decreased SS and VIP (P<0.01). The recurrence rate in the observation group was 34.29% (12/35),obviously lower than 61.90% (13/21) in the control group (χ2=5.051,P<0.05). The total effective rates in the observation group and the control group were 95.08% (58/61) and 81.67% (49/60) , implying a significant difference (χ2=5.321,P<0.01).ConclusionFor the elderly with primary constipation due to Qi-Yin deficiency, oral administration of modified Zhizhutang helps to regulate gastrointestinal neurotransmitters,improve gastrointestinal motility and gastrointestinal emptying rate, alleviate the symptoms of constipation, restore spontaneous bowel movements, improve their quality of life, and lower the recurrence rate. Its clinical effects were better than those of Qihuang Tongmi soft capsule.
Abstract:ObjectiveTo re-evaluate the intervention effect of Kuijietong(KJT) on ulcerative colitis(UC).MethodSixty patients with mild-to-moderate UC in the active stage were enrolled and randomized into a KJT group (n=30) and a sulfasalazine (SASP) group (n=30). Patients in the KJT group were treated with KJT granules, one bag divided in two daily doses, once in the morning and once in the evening, while those in the SASP group received SASP, 1 g per time, four times per day. Then the clinical efficacy was evaluated.ResultAccording to the modified Mayo score,the clinical remission rates of the KJT group and SASP group were determined to be 46.7% (14/30)and 40% (12/30),exhibiting no significant difference between the two groups (P>0.05). The clinical effective rate of the KJT group was 83.3% (25/30),which was better than 60% (18/30) of the SASP group (P<0.05). The mucosal healing rate in the KJT group was 36.7% (11/30), not significantly different from 30% (9/30) in the SASP group. In the alleviation of UC symptoms,the score of large intestine dampness heat syndrome in the KJT group was remarkably better than that in the SASP group (P<0.05),but there was no significant difference in inflammatory bowel disease questionnaire (IBDQ) score between the two groups. In terms of physical and chemical indexes,serum erythrocyte sedimentation rate (ESR) in the KJT group after intervention was lower than that in the SASP group (P<0.05),whereas the interleukin-10 (IL-10) level was higher(P<0.05). The comparison between the two groups revealed no significant difference in C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), CD4+ T cells and regulatory T (Treg) cells after intervention. During the intervention,no obvious adverse reactions were found in the two groups,indicating good safety.ConclusionKJT is not inferior to SASP in relieving mild-to-moderate UC in the active stage.
Abstract:ObjectiveTo observe the clinical efficacy of modified Huqianwan combined with auricular acupressure with seeds on girls with central precocious puberty (CPP) due to Yin deficiency and fire hyperactivity.MethodA total of 150 cases were randomly divided into three groups (A group,B group, and C group),with 50 cases in each group. The cases received modified Huqianwan,Zhibo Dihuangwan,and modified Huqianwan combined with auricular acupressure with seeds, respectively, for 12 weeks. The traditional Chinese medicine (TCM) symptoms,bone age difference/chronological age difference (ΔBA/ΔCA), predicted adult height (PAH),ovarian volume,uterine volume,breast nodule index,serum sex hormones [follicle-stimulating hormone (FSH),luteinizing hormone (LH),and estradiol (E2)],and serum kisspeptin-1 (KISS-1)/G protein-coupled receptor 54 (GPR54)/hypothalamic gonadotropin-releasing hormone (GnRH) signaling pathway of the three groups were observed before and after treatment. The clinical efficacy and safety of the three groups were evaluated.ResultThe total effective rate was 97.9% in the C group, which was higher than 81.3% in the A group (χ2=4.516,P<0.05) and 71.4% in the B group (χ2=5.162,P<0.05). Compared with the results in the A group after treatment,TCM syndrome,ΔBA/ΔCA,E2,LH,KISS-1,GPR54,and GnRH decreased(P<0.05),and ovarian volume,uterine volume,and breast nodule index were reduced (P<0.05),but PAH increased in the C group (P<0.05). Compared with the B group after treatment,the C group showed decreased TCM syndrome,ΔBA/ΔCA, FSH,E2,LH,KISS-1,GPR54,and GnRH (P<0.05),reduced ovarian volume,uterine volume,and breast nodule index (P<0.05),and increased PAH(P<0.05). During the study,there was one case of anorexia in the A group,and no obvious adverse reactions were found in the B group and the C group.ConclusionModified Huqianwan combined with auricular acupressure with seeds can significantly increase the PAH of girls with CPP due to Yin deficiency and fire hyperactivity and improve the serum sex hormones and the KISS-1/GPR54/GnRH signaling pathway.
Keywords:modified Huqianwan;auricular point pressing with bean;central precocious puberty;Yin deficiency and fire hyperactivity
Abstract:ObjectiveTo discuss the clinical efficacy of Xuesusheng granule for cancer related fatigue (CRF) deficiency of both Qi and blood in clinical practice, and to investigate its effect on immune function and quality of life.MethodAccording to random number table method, 118 patients were divided into observation group (59 cases) and control group (59 cases). Patients in control group got Ejiao Huangqi oral liquid, 10-20 mL/time, 2 times/day. Patients in observation group got Xuesusheng granule, 10 g/time, 3 times/day. The treatment was continued for 8 weeks in both groups. Before and after treatment, scores of cancer fatigue scale (CFS), functional assessment of cancer therapy (FACT) and Qi and blood deficiency syndrome were graded, and degree of fatigue was discussed with brief fatigue inventory scale-Chinese (BFI-C). Levels of white blood cell count (WBC), neutrophil count (NEU), platelet count (PLT), red blood cell count (RBC), and hemoglobin (Hb) were compared before and after treatment.T lymphocyte subsets of (CD3+, CD4+, CD8+) levels, natural killer (NK) cells, interleukin-1(IL-1), IL-6, IL-8 and tumor necrosis factor -α (TNF -α) were also detected, and the safety was evaluated.ResultAfter treatment the scores of each factor in CFS scale and the total score of CFS in the observation group were lower than those in the control group (P<0.01), and the degree of fatigue was lower than that in control group (Z=2.101, P<0.05). The total score of FACT and the scores of four dimensions of physiology, society/family, emotion and function in the observation group were lower than those in the control group (P<0.01). Levels of WBC, NEU, RBC and Hb in the observation group were higher than those in the control group (P<0.01). Scores of BFI-C, deficiency of Qi and blood were lower than those in control group (P<0.01). Levels of NK, CD3+, CD4+ and CD4+/CD8+ in the observation group were all higher than those in the control group (P<0.01), while the IL-1, CD8+, IL-6, IL-8 and TNF-α levels in observation group were lower than those in the control group (P<0.01). Not observed, there was no adverse reaction related to Xususheng granule.ConclusionXuesusheng granule can improve the symptoms of fatigue, reduce the degree of fatigue, improve the quality of life, reduce the degree of bone marrow suppression, improve immune function, and regulate immune inflammatory factors with high clinical efficacy in the use for patients with CRF and deficiency of Qi and blood.
Keywords:cancer related fatigue;syndrome of deficiency of Qi and blood;Xuesushen granule;quality of life;myelosuppression;immune function;inflammatory factors
Abstract:ObjectiveTo establish an ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) for determining the plasma concentrations of 10 active ingredients in Wujiwan at different time points after oral administration, and to compare the pharmacokinetic characteristics between normal rats and rats with chronic visceral hypersensitive irritable bowel syndrome (CVH-IBS).MethodCVH-IBS rat model was prepared by the neonatal rat colon percutaneous transluminal coronary angioplasty (PTCA) balloon stimulation method. After intragastric administration of Wujiwan (0.245 g·kg-1), blood was collected from the jugular vein at different time points, and the plasma concentrations of 10 active ingredients (berberine hydrochloride, palmatine hydrochloride, coptisine hydrochloride, jatrorrhizine hydrochloride, epiberberine, dihydroberberine, evodiamine, evodine, paeoniflorin, albiflorin) in Wujiwan was detected simultaneously by UPLC-MS/MS, the pharmacokinetic parameters of each component in normal rats and CVH-IBS rats were calculated.ResultThe established UPLC-MS/MS could sensitively and accurately detect the plasma concentrations of 10 active ingredients of Wujiwan in rats. Compared with the normal group, the absorption rates of these 10 active ingredients of Wujiwan in the blood of CVH-IBS rats all decreased to a certain extent, and the peak time (tmax) was prolonged. Among them, the tmax of berberine hydrochloride and jatrorrhizine hydrochloride were significantly prolonged from 54 minute and 39 minute to 90 minute, respectively (P<0.05, P<0.01). Area under the plasma concentration-time curve (AUC0-t) of each component increased, and evodiamine and paeoniflorin were significantly different (P<0.05, P<0.01). The clearance rates (CL/F) of these 10 active ingredients were all decreased, among which berberine hydrochloride, palmatine hydrochloride and evodiamine had significant differences (P<0.05, P<0.01).ConclusionThere are significant differences in the pharmacokinetic behavior of the active ingredients in Wujiwan between normal rats and CVH-IBS rats, which may be related to the destruction of microstructure of intestinal epithelial cells and the change of activity of liver enzymes under the pathological state of IBS.
Keywords:Wujiwan;irritable bowel syndrome (IBS);ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS);neonatal rat colon balloon stimulation method;pharmacokinetics;chronic visceral hypersensitivity;alkaloids
Abstract:ObjectiveTo explore the effect of different extracts of Thlaspi Herba on the gut microbiota of hyperuricemia mice, and to reveal the substance basis and mechanism of its hypouricemic activity.MethodEighty-eight male Kunming mice were divided into 11 groups, including blank group, model group, allopurinol group, high and low dose groups of petroleum ether extract, high and low dose groups of ethyl acetate extract, high and low dose groups of n-butanol extract, high and low dose groups of total flavonoids extract. Mice in the blank group were given 0.5% sodium carboxymethylcellulose by gavage, and the other groups were given oteracil potassium (500 mg·kg-1) by gavage to duplicate the hyperuricemia model. After modeling for several hours, the blank group and the model group were given distilled water by gavage, while mice in the allopurinol group were given allopurinol suspension (50 mg·kg-1), and mice in each treatment group were given high and low doses of corresponding extract (5, 2.5 g·kg-1). The serum uric acid (SUA) level and xanthine oxidase (XOD) activity were measured after 14 days. Fresh feces were collected for 16S rDNA sequencing.ResultCompared with the blank group, SUA level and XOD activity of model group were significantly increased (P<0.05). Compared with the model group, SUA level and XOD activity of the allopurinol group were significantly decreased (P<0.01). After intervention, SUA level were significantly decreased (P<0.05, P<0.01), except for high dose and low dose groups of petroleum ether extract and low dose group of total flavonoids extract, XOD activity was significantly inhibited in low dose group of petroleum ether extract, high dose group of total flavonoids extract, and high and low dose groups of n-butanol extract (P<0.05, P<0.01). The high dose group of total flavonoids extract was the most significant. The results of flora sequencing showed that α diversity and abundance of the model group changed significantly, and Bacteroidetes, Firmicutes and Lactobacillaceae were significantly correlated with XOD activity. After intervention, the operational taxonomic unit (OTU), ACE, Chao1 and Shannon indexes of the high and low dose groups of total flavonoids extract were significantly increased (P<0.05, P<0.01). Relative abundance of Bacteroidetes in low dose group of ethyl acetate extract, high dose group of total flavonoids extract, and high and low dose groups of n-butanol extract was significantly decreased (P<0.01), and the relative abundance of Firmicutes was significantly increased (P<0.01). The relative abundance of Lactobacillaceae in low dose group of n-butanol extract and high dose group of total flavonoids extract was significantly increased (P<0.01).ConclusionThe effective part of Thlaspi Herba for reducing uric acid is mainly flavonoids, the improvement of SUA level and XOD activity by affecting gut microbiota such as Lactobacillaceae, Bacteroidetes and Firmicutes, may be one of its mechanisms.
Abstract:ObjectiveTo study the effects of 24-epibrassinolide (EBR) at different concentrations on seed germination, seedling growth and key enzyme activities for flavonoid biosynthesis in Astragalus membranaceus var. mongholicus under polyethylene glycol (PEG)-induced drought stress, in order to provide theoretical reference for standardizing the drought-resistant cultivation techniques of A. membranaceus var. mongholicus.MethodThe seeds of A. membranaceus var. mongholicus were soaked in EBR solution at different concentrations(0.001, 0.01, 0.1, 1, 10 μmol·L-1) followed by foliar spraying to explore their effects on seed germination, seedling growth, photosynthesis, chlorophyll and malondialdehyde (MDA) contents, and key enzyme activities for flavonoid biosynthesis under drought stress induced by 20% polyethylene glycol (PEG)-6000.ResultCompared with the control(CK)group, PEG-induced drought stress led to an obvious decrease in germination potential, germination rate, germination index, vigor index, relative germination rate, plant height, root length, aboveground and root dry weight, net photosynthetic rate (Pn), stomata conductance (Gs), transpiration rate (Tr), chlorophyll contents and chlorophyll a/b ratio, while a significant increase in intercellular carbon dioxide concentration (Gi), MDA, flavonoids contents, and key enzyme activities for flavonoid biosynthesis such as phenylalanine ammonia lyase (PAL). The treatment with exogenous EBR solution at the suitable concentration significantly enhanced the adaptation of A. membranaceus var. mongholicus seeds and seedlings to PEG-induced drought stress, manifested as significantly elevated germination potential, germination rate, germination index, vigor index, relative germination rate, plant height, root length, aboveground and root dry weight, Pn, Gs, Tr, chlorophyll a/b ratio, chlorophyll and flavonoids contents and key enzyme activities for flavonoid biosynthesis like PAL while lowered Ci and MDA contents. The optimal concentration of EBR solution was 0.1 μmol·L-1.ConclusionExogenous EBR solution at the suitable concentration ameliorates the inhibitory effect of 20% PEG stress against seed germination and seedling growth of A. membranaceus var. mongholicus, reduces the oxidative damage in leaves, and improves the stress resistance to a certain extent by up-regulating the key enzyme activities and promoting flavonoid synthesis.
Abstract:ObjectiveTo investigate the percutaneous permeability of sinomenine hydrochloride (SNH) and optimize the parameters of electroporation to achieve the best permeation enhancing effect on SNH.MethodThe percutaneous permeability of SNH and the enhancement effect of electroporation were studied by in vitro diffusion cell method, and the enhancement effect of electroporation was further evaluated by in vivo study in mice.ResultUnder steady-state condition, the permeation rates of SNH in stripped skin and intact skin of hairless mice were (385.81±12.88), (0.88±0.20) μg·cm-2·h-1, respectively. The permeation rate in stripped skin was 438 times higher than that in intact skin. The results of percutaneous permeation kinetics analysis showed that the solubility and diffusion coefficient of SNH in stratum corneum were relatively low, which were (70.82±9.63)×103 g·m-3 and (3.07±1.52)×10-14 cm2·s-1, respectively. Under the optimized electroporation conditions (voltage of 72 V, time of 60 min), the 24 h cumulative permeation amount of SNH through skin of mice was (10 008.39±1 961.57) μg·cm-2, and the steady-state permeation rate was (456.01±51.26) μg·cm-2·h-1, which were 5.4 times and 5.1 times higher than those of blank group, respectively. In vivo studies in mice showed that the contents of SNH in skin and muscle of electroporation group were 2.0 times and 1.5 times higher than those of blank group.ConclusionThe low solubility and low diffusion coefficient of SNH in the stratum corneum are the main factors hindering the percutaneous permeation of SNH. Electroporation can significantly increase the percutaneous permeation of SNH and its retention in skin and muscle of mice.
Abstract:ObjectiveTo rapidly identify the chemical constituents of Chaishi Tuire granules by ultra-performance liquid chromatography-electrospray/quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS).MethodChromatographic separation was conducted on a Phenomenex® Luna omega C18 column (2.1 mm×100 mm, 1.6 μm) with 0.1% formic acid aqueous solution (A)-acetonitrile (B) as the mobile phases for gradient elution (0-20 min, 5%-40%B; 20-40 min, 40%-95%B; 40-43 min, 95%B), the flow rate was set at 0.3 mL·min-1. MS data were collected in positive and negative ion modes, the scanning range was m/z 150-1 500 and electrospray ionization (ESI) was employed. The chemical constituents of Chaishi Tuire granules were identified by comparing with the retention time and the mass data of the reference substances, as well as the accurate mass, MS/MS fragment ions, mass spectrometry databases (PubChem, MassBank, ChemicalBook and others) and related literature.ResultA total of 85 chemical constituents were identified, including 28 flavonoids, 24 phenylpropanoids, 11 terpenoids, 10 alkaloids, 4 quinones, and 8 others. Among them, 19 constituents derived from Lonicerae Japonicae Flos, 14 constituents derived from Scutellariae Radix, 10 constituents derived from Isatidis Radix, 9 constituents derived from Taraxaci Herba, 9 constituents derived from Forsythiae Fructus, 4 constituents derived from Bupleuri Radix, 4 constituents derived from Anemarrhenae Rhizoma, and 4 constituents derived from Rhei Radix et Rhizoma.ConclusionChaishi Tuire granules is rich in phytochemicals, which are derived from many of traditional Chinese medicines. This study can lay a foundation for the quality control, material basis and in vivo metabolic analysis of this preparation.
Keywords:Chaishi Tuire granules;ultra-performance liquid chromatography-electrospray/quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS);flavonoids;phenylpropanoids;terpenoids;alkaloids;fragmentation pattern
Abstract:ObjectiveTo analyze the functions and indications, formulation, dosage form and medication characteristics of Chinese patent medicines in the 2020 edition of Chinese Pharmacopoeia (part Ⅰ) for treating cough of children, and to provide ideas for the clinical rational application and provide reference for the research and development of new cough medicines for children.MethodThe name, dosage form, formulation, functions and indications, usage and dosage, and other information of Chinese patent medicines for cough were collected from the 2020 edition of Chinese Pharmacopoeia (part Ⅰ), then relevant information was input into Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine v2.0, and data analysis and mining were carried out through the analysis module of prescription medication rule, VOSviewer 1.6.14 was used to make drug clustering network view of Chinese patent medicines for the treatment of exogenous wind cold, exogenous wind heat and phlegm heat cough.ResultIn the 2020 edition of Chinese Pharmacopoeia (part Ⅰ), a total of 75 kinds of Chinese patent medicines for treating cough of children were collected, including 34 kinds of Chinese patent medicines for adults and children, 41 kinds of Chinese patent medicines for children only. There were 7 types of traditional Chinese medicine syndromes, such as wind-cold attacking the lung, wind-heat invading the lung and phlegm-heat obstructing the lung. There were 45 Chinese patent medicines for treating exogenous cough, accounting for 60%, among which 35 kinds were used for exogenous wind-heat cough and 10 kinds were used for wind-cold cough. There were 30 kinds of Chinese patent medicines for treating internal injury cough, including 19 kinds of medicines for phlegm heat obstructing the lung, 4 kinds of medicines for phlegm dampness containing the lung and phlegm food stagnation, 2 kinds of medicines for Yin-deficiency lung heat, 1 kind of medicine for the lung and spleen Qi-deficiency. The formulation analysis showed that Glycyrrhizae Radix et Rhizoma, Platycodonis Radix, Scutellariae Radix, Armeniacae Semen Amarum and Citri Reticulatae Pericarpium appeared frequently, which were mainly cold, bitter and sweet herbs, mainly belonged to the lung and stomach meridians. According to the analysis of administration and dosage forms, 71 kinds of Chinese patent medicines were administered through gastrointestinal tract, including 20 kinds of granules, 15 kinds of oral liquids, others included syrups, pills, capsules, tablets, powers, etc. Only 2 suppositories and 2 injections were administered by nongastrointestinal tract. The usage and dosage of most Chinese patent medicines were not clear.ConclusionIn the 2020 edition of Chinese Pharmacopoeia (part Ⅰ), the main syndromes of Chinese patent medicines for cough of children are exogenous wind-heat and phlegm-heat obstruction in the lung. Most of the Chinese medicines are cold, bitter and sweet, and their meridians are mainly lung and stomach meridians. Scutellariae Radix, Lonicerae Japonicae Flos and Forsythiae Fructus are the most common medicines of exogenous wind heat syndrome. Perillae Folium, Citri Reticulatae Pericarpium and Ephedrae Herba are the most common medicines of exogenous wind cold syndrome. Meanwhile, Scutellariae Radix, Platycodonis Radix and Armeniacae Semen Amarum are the most common medicines of phlegm heat obstructing the lung syndrome. At present, the dosage forms of Chinese patent medicines used for treating cough of children are too few and the dosage labeling is not comprehensive, so it is necessary to further strengthen the research and development of new Chinese medicines suitable for characteristics of children.
Keywords:Chinese Pharmacopoeia;children;cough;Chinese patent medicines;medication rule;medicinal properties;dosage forms
Abstract:ObjectiveThis study explores the key core targets of Guishenwan in the treatment of thin endometrium and related signaling pathways through the method of network pharmacology,and further uses animal experiments to verify the obtained targets and verify that Guishenwan are effective for thin endometrium.MethodTraditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) was used to retrieve the effective chemical components,active component targets and target abbreviations of the eight Chinese medicines in Guishenwan,the GeneCards database and Online Mendelian Inheritance in Man(OMIM)database were used to retrieve thin endometrial related targets gene.Use Wayne software to take the intersection of the drug target of Guishenwan and the disease target of the thin endometrium,and import the intersection target into the STRING database and Cytoscape 3.7.2 software for visual analysis to obtain the "drug-disease" protein protein interaction(PPI) network, then input the intersection target into Enrichr database and DAVID database for gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis. Using the obtained possible core and key targets as the theoretical basis,a thin endometrial model in rats was established. After Guishenwan and estrogen intervention for 21 days,the endometrial thickness of rats was observed by hematoxylin-eosin staining(HE) staining. Western blot and quantitative real time polymerase chain reaction(Real-time PCR) detect the protein and mRNA expression levels of the four core key targets of epidermal growth factor receptor(EGFR),matrix metalloproteinase 9(MMP9),interleukin-1beita(IL-1β) and mitogen activated protein kinase 14(MAPK14).ResultThe Venn software obtained 130 intersection targets in total, imported 130 intersection targets into the STRING database and Cytoscape database,and obtained a protein interaction network diagram including 33 nodes and 107 edges. DAVID 6.8 database for GO analysis. The function annotation analysis involving 167 biological processes(BP),22 cell components(CC),39 molecular functions(MF). DAVID 6.8 database for KEGG enrichment analysis, and thin endometrium related A total of 34 pathways. Western blot and Real-time PCR were used to analyze the expression results of EGFR,MMP9,IL-1β,MAPK14 protein and genes in the endometrial tissues of the 6 groups of rats. Guishenwan can enhance the expression of EGFR,MMP9,IL-1β,MAPK14 protein and mRNA on the thin endometrium.ConclusionAccording to the theoretical analysis of network pharmacology and the results of animal experiments,it is found that Guishenwan can effectively improve the related indicators of thin endometrium,and promote the expression of EGFR,MMP9,IL-1β,MAPK14 protein and genes. The intimal tissue proliferates and improves the symptoms of thin intima.
Keywords:Guishenwan;thin endometrium;network pharmacology;mechanism of action
Abstract:As a Chinese saying goes, "good Chinese medicinal material makes good medicine", the traceability of Chinese medicinal materials is closely related to the high-quality development of Chinese medicine industry. This article intends to analyze the "new era of smarter food safety blueprint" and Proposed Rule for Food Traceability related to food traceability of the U.S. Food and Drug Administration (FDA), including food traceability list, critical tracking events, and examples of exemptions. By studying the process and consideration of the construction of the FDA food traceability system, as well as the specific traceability requirements of fresh herbs, vegetables and fruits in the FDA food traceability system that are related to Chinese medicinal materials, on the one hand, it provides research materials for the communication and discussion of Chinese medicinal materials traceability, on the other hand, it also provides reference for the construction of food traceability system in China. To sort out the idea of constructing a traceability system for Chinese medicinal materials suitable for the current development status of traditional Chinese medicine, the contents to be considered are as follows:①To determine the traceability list of Chinese medicinal materials based on the risk control of Chinese medicinal materials. ②Determine the critical tracking links and information in the Chinese medicinal materials industry chain, and standardize the traceability information of Chinese medicinal materials. ③The quality information of Chinese medicinal materials should be reflected in the traceability of Chinese medicinal materials. ④Within the scope of the traceability list of Chinese medicinal materials, enterprises should be encouraged to make voluntary traceability by selecting Chinese medicinal materials with a good foundation of traceability work.
Keywords:Chinese medicinal materials;food;digital technology;traceability system of whole process;modern industrial system;critical tracking link;critical tracking information
Abstract:Gegen Qinliantang, a classic traditional Chinese medicine(TCM) compound, has been verified in modern research to possess various pharmacological effects such as anti-inflammation,anti-oxidative stress,protecting intestinal mucosal barrier, and regulating intestinal flora and immune response. Ulcerative colitis (UC) is a chronic idiopathic inflammatory disease involving the colorectal mucosa, which mainly results from genetic susceptibility, intestinal mucosal barrier damage, abnormal immune response, intestinal flora disturbance, and bile acid metabolism disorders. By reviewing the literature published in recent years, this paper sorted out the relevant pathways and mechanisms involved in the treatment of UC by Gegen Qinliantang to provide ideas for further clinical and basic research. This literature review uncovered that Gegen Qinliantang exerted the therapeutic effects against UC mainly via interleukin-6(IL-6)/Janus tyrosine kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3) signaling pathway, Toll like receptor 4(TLR4)/nuclear transcription factor-κB(NF-κB) signaling pathway,Notch signaling pathway, and matrix metalloproteinase-9(MMP-9)/p38 mitogen-activated protein kinase(p38 MAPK) signaling pathway. Gegen Qinliantang regulates the intercellular molecular transmission in multiple pathways to protect the intestinal mucosal barrier, adjust the immune response and anti-oxidative stress, and relieve UC, demonstrating the multi-target, multi-mechanism, and multi-pathway advantages of TCM compounds.
Abstract:The tumor incidence goes up with every passing year. Chemotherapy, as one of the main treatment methods, is faced with a major challenge of drug resistance in clinical practice. Tumor-associated macrophages (TAMs) are the key factors inducing chemotherapy resistance of tumors. TAMs are inflammatory cells with the largest number in the tumor microenvironment, which are widely distributed in such epithelial tissues as large intestine and stomach in the immune environment and closely associated with multiple common cancers like breast cancer and colorectal cancer. TAMs can be divided into two phenotypes, with M2-polarized TAMs into the tumor-promoting phenotype that affects the oncogenesis and progression and promotes drug resistance via immune escape, angiogenesis and other ways. At the same time, the frequently utilized chemotherapeutic agents will increase the recruitment of TAMs and trigger the secretion of cytokines, leading to the excessive polarization of macrophages to M2 type, followed by tumor drug resistance. The molecular mechanism of chemotherapy resistance is complex, which is becoming an urgent problem in the field of chemotherapy. Traditional Chinese medicine (TCM) has exhibited unique advantages in resisting tumor drug resistance. It has been proved efficient and safe in improving tumor microenvironment and regulating TAMs by acting on multiple targets via multiple ways, thus adjusting tumor progression and improving drug resistance. Based on related articles published in recent years, this paper reviewed the drug resistance-promoting effect of TAMs via regulating the immune microenvironment and interacting with tumor stem cells and the driving effect of chemotherapeutic agents on drug resistance to figure out the role of TAMs in chemotherapy resistance. Besides, it summarized the mechanisms of TCM in regulating related cytokines, proteins, activity, and the polarization direction of TAMs to expound the effective components of TCM in the intervention of drug resistance. The aim of this paper was to provide reference for further research on the biological mechanism of chemotherapy resistance and its targeted intervention with TCM.
Keywords:tumor-associated macrophages (TAMs);chemotherapy resistance;traditional Chinese medicine (TCM) treatment
Abstract:Shengxiantang, one of the classical traditional Chinese medicine (TCM) prescriptions, was developed by ZHANG Xi-chun, a well-known doctor in the Republic of China, to treat the syndrome of pectoral Qi sinking. Due to the unique herbal composition, this prescription has yielded remarkable outcomes, and it is still widely used for dealing with diseases of various systems. This paper reviewed the clinical research concerning Shengxiantang in order to provide reference for its clinical application. Following the retrieval of related articles published in the past ten years from China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform, Chongqing Weipu Database for Chinese Technical Periodicals (VIP), and Chinese Biomedical Literature Service System (SinoMed) with the Chinese key words of “Shengxiantang (升陷汤)”, “sinking of pectoral Qi (大气下陷)”, “replenishing Qi and raising the sinking (益气升陷)”, and“pectoral Qi theory (大气理论)”, this paper collated the clinical application experience of Shengxiantang. To be specific, the corresponding etiology and pathogenesis, interpretations of Shengxiantang, as well as its indications, application experience and precautions were summarized to provide a basis for its further research and application. Based on the collected more than 50 representative articles, it was found that Shengxiantang has been extensively utilized for treating complex and critical diseases of various systems, especially the circulatory and respiratory diseases and organ prolapse. The research on the clinical application of Shengxiantang has deepened, exhibiting a promising development trend, which will provide new ideas for its clinical application.
Keywords:Shengxiantang;syndrome of pectoral Qi sinking;replenishing Qi and raising the sinking;clinical application;review
Abstract:Volatile oil is widely distributed in Chinese medicinal materials with complex chemical components. The main components are terpenes, aromatics, aliphatics, and nitrogen and sulfur containing. It has a variety of pharmacological activities. Such as antibacterial, anti-inflammatory, analgesic, anti-tumor, anti-oxidant, anti-aging and so on. It is widely used in medical and health care, agricultural efficiency enhancement, and daily products. In recent years, there have been a large number of studies on the pain relief of traditional Chinese medicine (TCM) essential oils, but there is no systematic generalization. The author found that the mechanism of TCM essential oils to exert analgesic effects mainly includes regulation of the central nervous system, anti-inflammatory and analgesic, antispasmodic and analgesic effects by consulting Chinese and foreign literatures in recent years, but the exact mechanism needs to be further verified. This article reviews the research progress of TCM essential oil pain relief from the aspects of pain classification, generation, analgesic mechanism and combination of other technologies, in order to provide reference for related research in the future.
Keywords:essential oils of traditional Chinese medicine;analgesia;aromatherapy;types of pain;analgesic mechanism
Abstract:Depression is a mental or affective disorder, whose occurrence is closely related to the physical, psychological, and social environmental factors. Clinically, patients with depression often present with significant and long-lasting blue mood, impaired thinking and cognitive functions, reduced mobility, and even self-harm and suicide attempts. The pathogenesis of depression is complex and diverse, and there mainly exist the neurotransmitter hypothesis, immunodeficiency hypothesis, hypothalamus-pituitary-adrenal (HPA) axis activation hypothesis, brain-derived neurotrophic factor (BDNF) hypothesis, and intestinal flora hypothesis. In western medicine, it is mostly treated with antidepressant drugs, but the resulting side effects and repeated attack cannot be ignored. Traditional Chinese medicine(TCM) has obvious advantages in dealing with depression. The advocates of treating "Qi", "phlegm", "stasis", and "five zang organs" have all been proved effective. Depression is characterized by a long course of disease and repeated attack, which coincides with the concept of "deficiency" in TCM. Tonifying deficiency is a method for treating depression based on TCM syndrome differentiation. Under the guidance of this principle, the formulated prescriptions will produce the desired effects. This article reviewed the related clinical and experimental studies on depression treatment via deficiency tonification in recent years, and summarized the corresponding prescriptions for benefiting Qi, replenishing blood, nourishing Yin, and tonifying yang, respectively, so as to enrich the TCM theory of depression and provide new ideas for its clinical treatment.
Keywords:depression;deficiency;treating from deficiency;benefiting Qi;replenishing blood;nourishing Yin;tonifying Yang;research progress
Abstract:Xinjiang is rich in resources of medicinal plants, which are widely used in ethnic medicines because of their remarkable pharmacological activities and safeguard lives and health of people of all ethnic groups in Xinjiang and local health services. The ethnic medicines in Xinjiang are harmonious but different and united in diversity. Besides, they also promote each other and develop together, constituting an inseparable and important part of traditional Chinese medicine in China. Among them, Uygur medicine, Kazak medicine, Kirgiz medicine, Tajik medicine, and Tatar medicine are predominant in regional characteristics, but the development of the ethnic medicine industry in Xinjiang is backward. In recent years, Xinjiang ethnic medicines have been reported to possess significant pharmacological activities, which deserve further development and investigation. However, the development of ethnic medicines in Xinjiang has been restricted severely due to the lack of effective inheritance of practice and experience, weak basic research, insufficient brand awareness, resource destruction, etc. This study analyzed the medicinal value, development status, and prospects of Xinjiang ethnic medicines and summarized the problems in the modernization of ethnic medicines in Xinjiang to provide some scientific references for the follow-up development and clinical application of Xinjiang ethnic medicines, aiming to promote the sustainable development of traditional ethnic medicines of China.
Abstract:Polygonati Rhizoma was first recorded in Mingyi Bielu(《名医别录》) as both medicine and food due to its high edible and medicinal values.However,there is no systematic textual research on its use as medicine and food. Polygonati Rhizoma and Polygonati Odorati Rhizoma show similar morphology,so they are often confused.Clematis apiifolia and Polygonatum odoratum are often regarded as the original plants of Polygonati Odorati Rhizoma. By reviewing the related records in Chinese herbal books of the past dynasties,this paper aimed to clarify the relationship between Polygonati Rhizoma,C.apiifolia and P.odoratum,trace the historical evolution of Polygonati Rhizoma as medicine and food from the name,harvesting and processing,property,flavor and meridian tropism,efficacy and indications,edible and medicinal records,and contraindications,and summarize its application in ethnic medicine,in order to inherit and develop the ethnic medicine. The findings demonstrated that the name of Polygonati Rhizoma and its synonyms were derived from its pictogram,function,and place of origin,containing a wealth of myth implication and Taoism thought.Polygonati Rhizoma,C.apiifolia and P.odoratum are three species,different from the conclusions in previous textual research. The seedlings,flowers,seeds,and rhizomes of Polygonati Rhizoma all could be used for edible and medicinal purposes,with its edible records exceeding the medicinal records. There were 22 medical books from the ethnic minority groups such as The Annals of National Medicine in Yunnan,Zhongguo Minzu Yaozhi Yao,and Dictionary of Chinese Ethnic Medicine recording the knowledge and application of 12 species in Polygonatum by 26 ethnic minorities.Due to their different living environments,lifestyles,and cultural customs,ethnic minorities have formed their unique medical cultures.
Keywords:Polygonati Rhizoma;medicine and food from the same source;edible and medicinal prescriptions;herbal textual research;ethnic usage