Mechanism of Danggui Sini Decoction(当归四逆汤)for Atherosclerosis Obliterans Based on Network Pharmacology and Molecular Docking

Objective:Based on network pharmacology and molecular docking,we want to explore the pharmacological effects and mechanism of Danggui Sini Decoction in the treatment of lower extremity arteriosclerosis obliterans.Methods:Using TCMSP database to search the main active components of Danggui Sini Decoction(当归四逆汤,DSD)and their related targets,Cytoscape3.8.1 software was used to construct a"component-disease-target"interaction network.Meanwhile,DAVID database was used to enrich the key target genes with GO and KEGG functions.And we used Auto Dock Tools 1.5.6 for molecular docking.Results:A total of 45 candidate active molecules and 250 potential target proteins related to ASO were screened.Key genes such as TNF,IL6,VEGFA,MMP9,IL1B,CCL2,CXCL8,ICAM1,VCAM1,and IL10 are mainly through TNF signaling pathway,HIF-1 signaling pathway,cytokine-cytokine receptor interaction,Toll-like receptor signaling pathway,NF-kappa B signaling pathway,MAPK signaling pathway and other biological pathways exert their effects.The results of molecular docking showed that 5UUI-sesamin,5UUI-paeoniflorin,4XCT-sesamin,and 4XCT-sesamin have extremely strong binding ability.Conclusion:Danggui Sini Decoction(当归四逆汤,DSD)can synergistically exert pharmacological effects through multiple components,multiple targets,and multiple pathways.On the basis of regulating immunity and inhibiting smooth muscle proliferation,it can prevent the occurrence and development of ASO.

Danggui Sini decoction treatment of refractory allergic cutaneous vasculitis:A case report

BACKGROUND Allergic cutaneous vasculitis(ACV)is a difficult disease to treat.At present,there is no effective treatment for this condition.Traditionally,immunosuppressants and hormones have been primarily used in its management,but the treatment effect is suboptimal,and it has several side effects.CASE SUMMARY We present the case of a 19-year-old woman who presented at our hospital with a four-year history of symmetric skin lesions mainly affecting her lower extremities.She had previously undergone treatment with prednisolone acetate,cetirizine hydrochloride,and loratadine tablets but had not experienced any relief in her condition.Thereafter,she was treated with oral traditional Chinese medicine.Her skin damage gradually improved within two months of treatment initiation.After six months,the skin ulcers had completely subsided.No evidence of skin ulcer recurrence was observed during the subsequent follow-up.This report presents the first case of a female patient who received oral Danggui Sini decoction for the treatment of ACV.CONCLUSION Danggui Sini decoction may be a promising oral treatment for ACV patients.

Effect of Sini Decoction plus Ginseng on COVID-19 based on network pharmacological analysis

Objective:To explore the effect of Sini Decoction plus Ginseng on COVID-19 based on network pharmacological analysis.Methods:TCMSP platform was used to search the compounds of Sini Decoction plus Ginseng with oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18,and the results were input into the UniProt database and converted into standard target names;Genecards and OMIM database were used to find COVID-19 target,and the intersection targets were obtained and Venn diagram was drawn.Cytoscape 3.7.2 was applied to make the network diagram of composition-disease-target;The interaction database platform-String was used to analyze the target protein interaction,and the data were input into the software of Cytoscape 3.7.2 to make the network diagram;David database was used to analyze the accumulation of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathways of drug-disease intersection targets.The component target pathway network was constructed by using Cytoscape 3.7.2.Results:Altogether,112 active components of Sini Decoction plus Ginseng were screened,corresponding to 234 targets.Besides,261 COVID-19 targets were obtained after screening,and the Venn map showed that 50 targets were intersected.Quercetin,kaempferol,naringenin,andβ-sitosterol were found to have higher moderate values shown by the component-disease-target network.The target proteins with high PPI network analysis value were IL-6,MAPK8,MAPK3,MAPK1,TP53,TNF,and CASP3.GO enrichment function analysis showed that 341 biological processes,33 cell components,and 50 molecular functions were involved,which showed inflammatory reaction,cell response to lipopolysaccharide,exogenous apoptosis signal pathway,positive regulation of RNA polymeraseⅡpromoter transcription,cytokine activity,chemokine activity,heme binding,and other biological processes.KEGG signaling pathway enrichment function analysisshowed that there were 112 signaling pathways.It included TNF signaling pathway,tuberculosis,influenza A,PI3K Akt signaling pathway,HIF-1 signaling pathway,and Toll-like receptor signaling pathway.According to the component-target-pathway network diagram,quercetin,kaempferol,naringenin,andβ-sitosterol in Sini Decoction plus Ginseng may act on IL-6,MAPK8,MAPK3,MAPK1,TP53,TNF,CASP3,and other targets through TNF signaling pathway,MAPK signaling pathway,Tolllike receptor signaling pathway,PI3K-Akt signaling pathway,and HIF-1 signaling pathway.Conclusion:Sini Decoction plus Ginseng can affect COVID-19 through multiple active components,multiple targets,and multiple pathways.

Dissecting the underlying pharmaceutical mechanism of Danggui Buxue decoction acting on idiopathic pulmonary fibrosis with network pharmacology

Backgroud:Danggui Buxue decoction(DBD),a classical prescription in traditional Chinese medicine,has been found to have protective effect on bleomycin-induced pulmonary fibrosis in rats by reducing alveolar inflammation and fibrosis.However,the biological activity of individual chemical components and mechanism of action of whole formula are not clear.Methods:Potential targets of active ingredients of DBD were collected through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and SymMap database.Target genes related to idiopathic pulmonary fibrosis were obtained from the Online Mendelian Inheritance in Man database,Therapeutic Targets Database and Gkb database.Then,the common targets were obtained by overlapping the potential targets of active ingredients in DBD and diseases related targets.The selected targets were subjected to Kyoto Encyclopedia of Genes and Genomes signaling pathway and Gene Ontology analysis,and the network map of active component-target-pathway was established using Cytoscape 3.7.1 software.The active components of DBD with most targets were selected for fibrosis-related marker verification.The mRNA and protein expression of fibrosis markers,α-smooth muscle actin,collagen 1 and fibronectin,were detected in TGF-β1-induced fibroblast cell line after treatment with the active components.Results:The 14 active ingredients,such as quercetin and kaempferol,were screened from DBD.It acts on 26 targets like estrogen receptor 2 and prostaglandin-endoperoxide synthase 2,and mainly involves 38 signaling pathways such as cell inflammation and autophagy.Kaempferol and quercetin are the two compounds with the highest network regulation,which can inhibit the transformation of fibroblasts into myofibroblasts and reduce the expression of fibrosis markersα-smooth muscle actin,collagen 1 and fibronectin.Conclusion:The integration mode of multi-component,multi-target,multi-channel and mechanism of DBD in the treatment of idiopathic pulmonary fibrosis are predicted by means of network pharmacology.Our study could indicate the direction of further anti-fibrotic mechanism research.

Danggui Niantong decoction ameliorates joint inflammation and cardiopulmonary injury in TNF-Tg mice

Objective:Rheumatoid arthritis(RA)is a common autoimmune disease characterized by multiple joint lesions and systemic complications.Danggui Niantong decoction(DGNTT)has been clinically used for RA treatment;however,its beneficial effect on cardiopulmonary complications has not been reported.Methods:Female tumor necrosis factor-transgenic(TNF-Tg)mice were used to evaluate the therapeutic effects of DGNTT on arthritis and cardiopulmonary complications.Methotrexate(MTX)served as a positive control.Histopathological assessment of the joint sections was performed using hematoxylin and eosin(HE),Alcian Blue/Orange G,and tartrate-resistant acid phosphatase staining.Bone mass was assessed by micro-computed tomography,inflammatory infiltrates in the heart and lungs were evaluated by HE staining,cardiopulmonary fibrotic injury was identified by Masson’s trichrome staining,and hypertrophy of mouse cardiomyocytes was measured by wheat germ agglutinin(WGA)staining.Results:DGNTT mitigated the inflammation of the ankle joint synovium,decreased the number of osteoclasts,and increased the area of cartilage and bone mass in TNF-Tg mice.In addition,DGNTT decreased the infiltration of inflammatory cells into the lung and heart tissues,accompanied by a reduction in cardiopulmonary fibrosis and myocardial cell hypertrophy in TNF-Tg mice.As a positive control drug,MTX attenuated the pathological changes in joints,but had no beneficial effect on cardiopulmonary inflammation and fibrosis in TNF-Tg mice.Conclusions:DGNTT improved joint lesions and alleviated cardiopulmonary complications in TNF-Tg mice.

Potential Mechanism of Danggui Buxue Decoction in Treating Iron Deficiency Anemia Based on Network Pharmacology and Molecular Docking Technology

[Objectives]To explore the potential mechanism of Danggui Buxue Decoction in treating the iron deficiency anemia(IDA)based on network pharmacology and molecular docking technology.[Methods]The active components and target proteins of Danggui Buxue Decoction were searched in databases such as TCMSP,OMIM,GeneCards,Drugbank,String,Metascape,etc.,and the target proteins shared with IDA were screened out,and the information about the signal pathways and biological functions of these target proteins was obtained.[Results]17 active components of Danggui Buxue Decoction and 24 potential targets for the treatment of IDA were obtained.With the aid of String database and Cytoscape software,the protein interaction network was obtained and the network topology analysis was performed.Four potential core targets with higher scores were obtained,namely F2,NOS2,NOS3,and PPARG.Using the Metascape database,GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the potential targets of Danggui Buxue Decoction in the treatment of IDA,and the important biological processes,cell composition,molecular functions and signal pathways related to the target were screened through the R language.The results show that biological processes are related to positive regulation of growth,cell composition is related to membrane microdomain,and molecular functions are related to oxidoreductase activity.The signal pathways involved are mainly AGE-RAGE signal pathway,TNF signal pathway and IL-17 signal pathway.Finally,the molecular docking results confirmed that the active components of Danggui Buxue Decoction have a good binding ability with the target.[Conclusions]Danggui Buxue Decoction treats the IDA through multiple components,multiple targets,multiple signal pathways,and multiple biological functions.

Study on the Molecular Mechanism of Danggui Buxue Decoction in Intervention of Perimenopausal Syndrome Based on Network Pharmacology and Molecular Docking

[Objectives]This study was conducted to explore the intervention mechanism of Danggui Buxue Decoction in perimenopausal syndrome based on network pharmacology and molecular docking.[Methods]The chemical components and targets of Danggui Buxue Decoction were acquired through the TCMSP database,and the main targets of perimenopausal syndrome were obtained through the GeneCards database.The component targets and disease targets were intersected,and combining with active components and Chinese herbs in the decoction,a traditional Chinese medicine-component-target network was constructed using Cytoscape 3.7.1 software.The STRING platform was employed for protein-protein interaction analysis.The DAVID analysis platform was used to conduct target GO and KEGG enrichment analysis,so as to predict the action mechanism Danggui Buxue Decoction.Finally,an active component-disease target-signal pathway network diagram was constructed.[Results]Twenty two components in Danggui Buxue Decoction related to perimenopausal syndrome and 120 corresponding targets were obtained,including active components such as 1,7-dihydroxy-3,9-dimquercetin and kaempferol,and key targets such as TNF,ESR1 and PPARG.The results of GO analysis and KEEG analysis indicated that Danggui Buxue Decoction might regulate the transcription of RNA polymerase II promoter,DNA templating,gene expression,signal transduction,hypoxia response and other biological processes by regulating multiple signal pathways such as chemical carcinogenesis-receptor activation,cancer pathways,lipid and atherosclerosis,tryptophan metabolism,malaria,steroid hormone biosynthesis and chemical carcinogenesis-DNA adduct.[Conclusions]Danggui Buxue Decoction intervenes in perimenopausal syndrome through multiple components,targets and pathways,providing a basis for elucidating the intervention mechanism of Danggui Buxue Decoction and expanding its clinical application.

Exploring the mechanism of Danggui Buxue Decoction in treating osteoporosis from vitamin D-axis

Osteoporosis is a common disease in the middle-aged and elderly population and is a global health problem that needs to be solved urgently.Vitamin D deficiency is closely related to osteoporosis,and it is of practical significance to prevent and treat kidney and strengthen bone therapy.A large number of literature have confirmed that qi and blood to represent Danggui Buxue Decoction has a reliable therapeutic effect on osteoporosis,but the mechanism is still unclear.Based on the previous work of the research group,it is believed that the vitamin D axis may be a potential target for TCM Bushen Recipe.Syndrome and vitamin D deficiency have similar pathological changes,so it is suggested that Danggui Buxue Decoction may regulate osteoporosis by regulating the vitamin d axis.

Effect of Danggui Buxue Decoction on the expression of VDR,CYP27B1,CYP19,and ERβin the ovary of osteoporosis rats

Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,FSH,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.The results of this experiment showed that compared with the control group,the serum FSH level of the model group was significantly increased,the Klotho level was decreased,the relative expression of ovarian tissue CYP27B1,CYP19,ERβmRNA and protein was significantly reduced,and the relative expression of vitamin D receptor mRNA and protein was significantly rise.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum FSH content decreased,Klotho content increased,ovarian CYP27B1,CYP19,ERβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Based on the effect of the classic ancient prescription Danggui Buxue decoction on the vitamin D system,combined with the results of this experiment,it is concluded that the classic ancient prescription Danggui Buxue decoction may affect the levels of CYP19,ERβ,follicle-stimulating hormone,and Klotho in the body by regulating the expression of ovarian VDR and CYP27B1.The specific mechanism of the treatment of ovarian injury in osteoporosis rats needs further study.Background:Investigate the effects of the classic ancient prescription Danggui Buxue decoction on the vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase and estrogen receptorβmRNA and protein expression in osteoporotic rats.Methods:The experiment was divided into 6 groups:control group,model group(retinoic acid 100 mg/kg/d),vitamin D group(calcitriol 30 ng/kg/d),the classic ancient prescription Danggui Buxue decoction high,middle and low dose group(7.2 g/kg/d,3.6 g/kg/d,1.8 g/kg/d).Enzyme-linked immunosorbent assay method was used to detect the effects of different doses of the classic ancient prescription Danggui Buxue decoction on serum follicle-stimulating hormone and Klotho levels in retinoic acid-induced osteoporosis rats;real-time fluorescence quantitative PCR was used to detect the classic ancient prescription Danggui Buxue decoction on retinoic acid effect of ovary vitamin D receptor,25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA expression in induced osteoporosis rats;Western-blot detection of the classic ancient prescription Danggui Buxue decoction on retinoic acid-induced osteoporosis ovary vitamin D receptor,CYP27B,estrogen synthesis aromatase,estrogen receptorβprotein The impact of expression.Results:Compared with the control group,the serum follicle-stimulating hormone level of the model group was significantly increased,the Klotho level was decreased,the relative expression levels of 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein in the ovarian tissue were significantly decreased,and the relative expression levels of vitamin D receptor mRNA and protein were significantly increased.After the intervention of the classic ancient prescription Danggui Buxue decoction and vitamin D,compared with the model group,the serum follicle-stimulating hormone content decreased,Klotho content increased,ovarian 25-hydroxyvitamin D-1αhydroxylase,estrogen synthesis aromatase,estrogen receptorβmRNA and protein expression significantly increased,ovarian vitamin D receptor mRNA and protein expression significantly decreased.Conclusion:The classic ancient prescription Danggui Buxue decoction may affect the levels of estrogen synthesis aromatase,estrogen receptorβ,follicle-stimulating hormone,and Klotho in the body through regulating the expression of ovarian vitamin D receptor and 25-hydroxyvitamin D-1αhydroxylase to achieve the therapeutic effect of retinoic acid-induced osteoporosis in rats.

A Medical Case of Acne Vulgaris Treated with Danggui Shaoyao Powder Combined with Sini Powder

Acne vulgaris is a chronic inflammatory skin disease damaging face,and the incidence of acne vulgaris is increasing year by year in China.Acne falls into the category of pimple in traditional Chinese medicine,and there are many dialectical understandings of acne in modern traditional Chinese medicine.Dr.He Aijuan has a unique diagnosis and treatment thinking in treating acne vulgaris(liver stagnation and spleen deficiency type)with traditional Chinese medicine in her clinical work for many years.Her application of Danggui Shaoyao Powder combined with Sini Powder has a remarkable effect in the clinical treatment process,so a clinical medical case is selected for discussion.

Danggui Buxue decoction promotes proliferation and differentiation of Caco-2 cells and antagonizes obesity by stimulating apolipoprotein-IV expression

Background:Chinese medicine has been proposed as a novel approach to the prevention of metabolic disorders such as obesity.Danggui Buxue decoction,a decoction prepared from Huangqi(Astragali Radix)and Danggui(Angelicae Sinensis Radix),has been used to nourish vitality and enhance blood circulation in traditional Chinese medicine.However,the effect of Danggui Buxue decoction on obesity is still primarily unknown.Methods:Cell proliferation,differentiation,and apolipoprotein-IV transcription were investigated to explore the function of Danggui Buxue decoction by methyl thiazolyl tetrazolium assay,alkaline phosphatase assay,and luciferase assay,respectively.Results:Danggui Buxue decoction promoted cell growth by up to 15%(P=0.034)and induced cell differentiation by up to 38%(P=0.006)at 0.3 mg/mL.Moreover,Danggui Buxue decoction enhanced the transcription of apolipoprotein-IV by 2.4 times(P=0.027),activating its promoter by 28.9%(P=0.031)at 0.3 mg/mL.In addition,Danggui Buxue decoction functioned in a dose-dependent manner.Conclusion:These results suggest that Danggui Buxue decoction promotes cell differentiation by enhancing apolipoprotein-IV transcription and alkaline phosphatase activity,and it may also have a potential anti-obesity effect because apolipoprotein-IV transcription is closely related to the reduction of food intake.

基于UHPLC-Q-Orbitrap/MS鉴定当归四逆颗粒化学成分

本研究运用超高效液相色谱-四极杆-静电场轨道阱质谱(UHPLC-Q-Orbitrap/MS)技术快速鉴定当归四逆颗粒的化学成分。采用SUPELCO C18色谱柱(100 mm×4.6 mm×2.7μm),以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,正、负离子模式下采集数据。根据精确质荷比和二级质谱碎片离子信息,结合相关文献快速鉴定当归四逆颗粒的成分,并对化合物的药味来源进行归属。结果表明,在正、负离子模式下,分别从当归四逆颗粒鉴定出34、38种化合物,其中包括20种黄酮类、15种有机酸类、11种皂苷类、9种萜类、5种苯酞类、1种苯丙素类、1种醛类、1种核苷类、1种木脂素类及8种其他类化合物。本研究可为阐释当归四逆颗粒的药效物质基础提供依据。

Research progress of Sini decoction in pharmacy, pharmacology and clinical application

Sini decoction originated from Zhang Zhongjing's Treatise on Cold Pathogenic and Miscellaneous Diseases(200-210 C.E.)in the Eastern Han Dynasty.It is a traditional and famous prescription in China.Its preparations mainly include Sini decoction,Sini decoction drop pills and Sini decoction suppositories.The main chemical components are diterpenoid alkaloids,gingerol,volatile oil,coumarin,polysaccharide,triterpene saponins,flavonoids,amino acids,and alkaloids.This product has a cardiovascular function,anti-shock,anti-diabetes complications,anti-tumor,anti-atherosclerosis,and effects on the nervous system.Modern clinical practice is widely used in cardiovascular,respiratory,digestive,neurological and gynecological diseases.This paper reviews the pharmaceutical research,pharmacological effects,safety evaluation and clinical application of Sini decoction,including the source and composition of Sini decoction,the origin and resource status of authentic medicinal materials of Sini decoction,the research on preparation form reform,chemical composition and quality control,to provide a reference for further research and clinical promotion of Sini decoction.

Comprehensive analysis of the potential biological mechanism of Danggui Buxue Decoction in the treatment of Thalassemia

Background:This study aimed to deeply explore the active components of Danggui Buxue Decoction(DBD)through systematic network pharmacology and molecular docking methods and to demonstrate its mechanism of regulating Thalassemia.Methods:Obtain and screen the active ingredients and targets of DBD from the TCMSP database.Use the GeneCards database to search for the gene corresponding to the target.The target protein-protein interaction(PPI)network was built using STRING database.A DBD-drug-Tha-target PPI network was established,and its core target network was analyzed using Cytoscape software.Then,enrichment analysis of DBD core targets was performed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG),and their binding activities were verified by molecular docking.Results:Finally,19 active ingredients and 504 targets were obtained from DBD,of which 98 targets were repeated with the related targets of Tha.The major genes include HSP90AA1,IL-6 and HRAS.Through the final analysis,we found that the main mechanisms of DBD in the treatment of thalassemia include increasing the expression of the γ-globin gene in red blood cells,inducing fetal hemoglobin and reducing the inflammatory response.Conclusion:The potential mechanism of DBD improving thalassemia can be preliminarily predicted by network pharmacology and molecular docking,which can provide some reference for clinical treatment.

Network pharmacology and experimental validation to reveal the pharmacological mechanisms of Sini decoction(四逆汤)against renal fibrosis

OBJECTIVE:To investigate the mechanism by which Sini decoction(四逆汤,SND)improves renal fibrosis(Rf)in rats based on transforming growth factor β1/Smad(TGF-β1/Smad)signaling pathway.METHODS:Network pharmacology was applied to obtain potentially involved signaling pathways in SND's improving effects on Rf.The targets of SND drug components and the targets of Rf were obtained by searching databases,such as the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCSMP)and GeenCard.The intersection targets of two searches were obtained and underwent signaling pathway analysis using a Venn diagram.Then experimental pharmacology was utilized to prove and investigate the effects of SND on target proteins in the TGF-β1/Smad signaling pathway.The Rf rat model was established by unilateral ureteral occlusion(UUO).The expression levels of transforming growth factor,matrix metalloproteinase-9(MMP-9),matrix metal protease-2(MMP-2),connective tissue growth factor(CTGF),and tissue inhibitor of metalloproteinase-1(TIMP-1)were determined by Masson staining of rat renal tissue,and immunohistochemical methods.The expression levels of Smad3,Smad2,and Smad7 in renal tissue were determined by Western blotting(WB).The mechanism of the improving effects of SND on Rf was investigated based on TGF-β1/Smad signaling pathway.RESULTS:A total of 12 drug components of Fuzi(Radix Aconiti Lateralis Preparata),5 drug components of Ganjiang(Rhizoma Zingiber),and 9 drug components of Gancao(Radix Glycy et Rhizoma)were obtained from the database search,and 207 shared targets were found.A total of 1063 Rf targets were found in the database search.According to the Venn diagram,in total,96 intersection targets were found in two database searches.The metabolic pathways involved included TGF-β signaling pathway,phosphatidylinositol-3-kinase/serine-threonine protein kinase signaling(PI3K/Akt)pathway,and hypoxia-inducible factor-1(HIF-1)signaling pathway.Masson staining analysis showed that compared with the model group,the renal interstitial collagen deposition levels in the SSN and SND groups were significantly lower(P<0.05).Immunohistochemical analysis,compared with the control group,the positive cell area expression levels of MMP-9/TIMP-1 and MMP-2/TIMP-1 in the kidney tissue of the model group were significantly decreased(P<0.05,P<0.01),and the positive cell area expression levels of CTGF and TGF-β1 were significantly increased(P<0.01).Compared with the model group,the positive cell area expression levels of MMP-9/TIMP-1 and MMP-2/TIMP-1 in the kidney tissue of the SSN and SND groups were significantly increased(P<0.05,P<0.01),and the positive cell area expression levels of CTGF and TGF-β1 in the kidney tissue were significantly decreased(P<0.05,P<0.01).WB results showed that the SSN group and the SND group could reduce the expression of Smad2 and Smad3(P<0.05)and increase the expression of Smad7(P<0.05).

UPLC-Q-Orbitrap-MS结合网络药理学探讨当归四逆汤治疗关节炎及痛经“异病同治”机制分析

目的基于超高效液相色谱串联四级杆-静电场轨道阱高分辨质谱技术(UPLC-Q-Orbitrap-MS)结合网络药理学探究当归四逆汤治疗类风湿性关节炎及原发性痛经“异病同治”的活性成分及作用机制。方法UPLC-Q-Orbitrap-MS技术对经典名方当归四逆汤的化学成分定性分析。采用中药系统药理学数据库与分析平台(TCMSP)、SwissTargetPrediction数据库对鉴别的化学成分预测靶点;采用Drugbank、OMIM、Genecard数据库收集与类风湿性关节炎、原发性痛经相关的靶点;利用Cytoscape3.7.2软件构建“活性成分—靶点—疾病”网络并分析关键成分和靶点;利用Metascape3.5在线对潜在靶点进行基因本体论功能注释(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路分析。结果共筛选出当归四逆汤治疗类风湿性关节炎及原发性痛经交集靶点96个,筛选出25个核心靶点,富集结果分析显示,990个涉及生物过程、73个涉及细胞组分、100个涉及分子功能。结论结果表明,当归四逆汤可能通过布雷非德菌素A、木犀草素-7-葡萄糖苷、咖啡酸、甘草素等成分作用于酪氨酸激酶(sarcoma gene,SRC)、信号转导子和转录激活子3(signal transducer and activator of transcription 3,STAT3)、表皮生长因子受体(Epidermal growth factor receptor,EGFR)、丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、磷酸肌醇3激酶的p110α催化亚基(phosphatidylinositol-4,5-bisphosphate 3-kinase,PIK3CA)、白介素-6(interleukin-6,IL-6)、花生四烯酸等信号通路发挥抗炎镇痛作用。

加味茵陈四逆汤对胆道闭锁肝纤维化幼鼠IL-17、IL-6与Th17及其特异性转录因子RORγt之间的影响

目的 探讨加味茵陈四逆汤对胆道闭锁(Biliary atresia, BA)肝纤维化幼鼠白介素-17(Interleukin-17,IL-17)、白介素-6(Interleukin-6,IL-6)与辅助性T细胞17(T helper cell 17,Th17)及其特异性转录因子维甲酸相关孤儿受体γt(RetinoicAcid-relatedOrphangammat, RORγt)之间的影响。方法 雄性SD大鼠幼鼠48只,随机分为假手术组、模型组、阳性对照组及加味茵陈四逆汤高、中、低剂量组,每组8只。建立幼鼠胆道结扎模型,5 d后进行灌胃给药。用药10 d后,计算肝脏指数;各组行苏木精-伊红染色法(Hematoxylin-eosin, HE)及Masson染色观察肝组织病理改变;采用酶联免疫吸附试验测定血清IL-17和IL-6水平;采用流式细胞仪测定Th17细胞占CD+4T细胞比例;采用实时荧光定量聚合酶链式反应(Polymerase Chain Reaction, PCR)法测定RORγt mRNA表达。结果 模型组及各给药组肝脏指数高于假手术组(P<0.05);各给药组肝脏指数低于模型组(P<0.05);各给药组肝纤维化程度较模型组减轻;模型组及各给药组IL-17、IL-6、Th17细胞占CD_(4)^(+)T细胞比例和RORγt mRNA表达高于假手术组(P<0.05);各给药组IL-17、IL-6、Th17细胞占CD_(4)^(+)T细胞比例和RORγt mRNA表达低于模型组(P<0.05);中剂量组和高剂量组IL-17、IL-6、Th17细胞占CD+4T细胞比例和RORγt mRNA表达低于熊去氧胆酸组和低剂量组(P<0.05)。结论 加味茵陈四逆汤可有效改善BA幼鼠肝纤维化程度,可能是通过降低血清IL-17和IL-6水平,从而降低Th17细胞水平及下调RORγt表达来发挥作用的。

Effect of the herbal medicine Danggui Sini plus Wuzhuyu Shengjiang Tang on erythrocyte deformability in normal subjects: a cross-over trial

OBJECTIVE: To show whether Danggui Sini plus Wuzhuyu Shengjiang Tang(DSWST) has any transient effect on erythrocyte deformability in normal subjects.METHODS: A total of 25 subjects [mean age(27.8± 1.8) years] was enrolled in this study. The study was designed as a cross-over trial in which the subjects took part for 2 d. On the first day, blood samples were collected at baseline and 1-2 h after administration of water, whereas, on the second day, instead of water, the subjects were administered DSWST after the baseline blood sampling. The blood samples collected at baseline and after the administration water or DSWST, were examined for erythrocyte deformability.RESULTS: The elongation index increased significantly after 2 h(P = 0.009) compared to the baseline after DSWST intake. However, after water intake, there was no significant differenceobserved. When comparing the percent change of erythrocyte deformability between DSWST and water, we found that after 2 h of administration,DSWST improved erythrocyte deformability significantly compared to water(P < 0.001).CONCLUSION: DSWST has a transient effect on erythrocyte deformability in normal subjects.

基于化学成分与药理作用评价当归-黄芪药对配方颗粒汤剂与传统汤剂的差异

目的评价当归-黄芪药对配方颗粒汤剂(DGD)与传统汤剂在化学成分和药理效应方面的差异,为合理应用该配方颗粒提供参考。方法以原料来源批次统一和不统一2种对比方式,设不同样品组,采用高效液相色谱(HPLC)法建立特征图谱,从特征图谱相似度、成分种类、指标成分含量、共有峰面积等角度对化学成分进行评价;采用失血性血虚模型小鼠评价药效。结果①DGD特征图谱与传统汤剂相似度较高(相似度>0.87);②共有色谱峰数目不一致,传统汤剂-自购饮片和传统汤剂-A厂饮片共有色谱峰各12个,A厂DGD共有色谱峰15个,B厂DGD共有色谱峰10个;③A厂DGD中阿魏酸、毛蕊异黄酮苷含量高于传统汤剂(P<0.05);B厂DGD与传统汤剂阿魏酸含量差异无统计学意义,但毛蕊异黄酮苷含量较传统汤剂低(P<0.05);④DGD共有峰面积总和与传统汤剂相比,自购传统汤剂、A厂传统汤剂及A厂配方颗粒、B厂配方颗粒中各成分含量相对比值分别为1.00、0.96、2.14、0.60;⑤DGD及传统汤剂均能明显促进失血性贫血模型小鼠血红蛋白(Hb)、红细胞(RBC)恢复(P<0.01);与模型对照组比较,除B厂DGD组外均差异有统计学意义(P<0.05);A厂DGD与传统汤剂差异无统计学意义,B厂DGD与传统汤剂差异有统计学意义(P<0.01)。结论无论原料来源批次是否一致,DGD与传统汤剂在化学成分上均存在一定差异;在药理作用上,来源于同一批次饮片制备的DGD与传统汤剂对改善失血性贫血药效相当,来源于不同批次饮片制备的DGD与传统汤剂在药效上存在一定差异;不同来源批次的饮片和不同制备工艺造成不同厂家配方颗粒存在质量差异,说明国家统一配方颗粒质量标准及制定相关过程规范具有必要性与紧迫性。

四逆汤中8种成分在大鼠体内的药动学研究

目的研究四逆汤中8种成分在大鼠体内的药动学特征。方法SD大鼠灌服四逆汤水煎液(4 g·kg^(-1)),不同时间点采血,LC-MS/MS法测定血药浓度,并计算药动学参数。结果乌头碱、新乌头碱、次乌头碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱、甘草苷、甘草酸和甘草次酸在相应浓度范围内线性关系良好(r>0.995),日内、日间RSD均<12%,达峰时间(T_(max))分别为(1.08±0.38)h、(1.08±0.38)h、(0.61±0.35)h、(1.32±0.76)h、(1.67±0.29)h、(0.25±0.12)h、(2.50±1.32)h和(10.67±2.31)h;消除半衰期(t_(1/2))分别为(2.68±0.78)h、(2.61±1.68)h、(5.56±3.15)h、(2.47±1.25)h、(9.12±4.89)h、(0.89±0.22)h、(10.50±6.84)h和(7.20±2.97)h;达峰浓度(C_(max))分别为(0.03±0.01)ng·mL^(-1)、(0.02±0.003)ng·mL^(-1)、(1.11±0.15)ng·mL^(-1)、(0.21±0.07)ng·mL^(-1)、(0.06±0.03)ng·mL^(-1)、(14.87±3.84)ng·mL^(-1)、(72.60±35.06)ng·mL^(-1)和(249.83±130.32)ng·mL^(-1);浓度-时间曲线下面积(AUC_(0-t))分别为(0.09±0.02)ng·h·mL^(-1)、(0.05±0.009)ng·h·mL^(-1)、(4.09±0.67)ng·h·mL^(-1)、(0.99±0.06)ng·h·mL^(-1)、(0.42±0.03)ng·h·mL^(-1)、(19.04±2.77)ng·h·mL^(-1)、(376.23±48.35)ng·h·mL^(-1)和(2726.65±1172.56)ng·h·mL^(-1)。结论本方法方便、快速,可用于四逆汤各成分的药动学研究。