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.

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.

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.

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.

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.

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.

Efficacy of Danggui Buxue decoction(当归补血汤)on diabetic nephropathy-induced renal fibrosis in rats and possible mechanism

OBJECTIVE:To observe the efficacy of Danggui Buxue decoction(当归补血汤,DBD) on diabetic nephropathyinduced renal fibrosis in rats,and to study the possible mechanism.METHODS:Sixty male Goto Kakizaki(GK) rats were randomly assigned to the model group,gliquidone group,astragaloside Ⅳ group,and high-,medium-and lowdoses DBD groups.After 8 weeks,changes in body weight,blood glucose,serum creatinine,serum urea nitrogen,and total cholesterol were observed.Changes in transforming growth factor-β1(TGF-β1),Smad3,and Smad5 pathways and the expression of the fibrosisrelated proteins collagen Ⅳ(col Ⅳ),α-smooth muscle actin(α-SMA),and vimentin were assessed.The degree of renal fibrosis was observed by immunohistochemistry and Mason staining.The expression of interleukin 6(IL-6),interleukin 10(IL-10),tumor necrosis factor(TNF-α),and C-reactive protein(CRP) in the kidneys was assessed using enzyme linked immunosorbent assay.RESULTS:Our experiments showed that DBD effectively reduced blood glucose,blood urea nitrogen,and creatinine levels after 8 weeks of administration,improved renal function in diabetic rats,alleviated renal fibrosis,and reduced the renal tissue levels of IL-6,IL-10,TNF-α,and CRP.Furthermore,DBD decreased the expression of TGF-β1,Smad3,col IV,α-SMA,and vimentin in renal tissues and increased the expression of Smad5.CONCLUSIONS:DBD ameliorates diabetic renal interstitial fibrosis by modulating the TGF-β1/Smads pathway.

Protection of Ultra-filtration Extract from Danggui Buxue Decoction(当归补血汤) on Oxidative Damage in Cardiomyocytes of Neonatal Rats and Its Mechanism

Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide （H2O2） and its potential mechanism. Methods： Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco＇s modified Eagle＇s medium low-glucose and Ham＇s F12 medium （1：1）, and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarurn po/ybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （Ml-r） assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase （LDH） and creatine kinase （CK） releases in the medium, superoxide dismutase （SOD） activities, and intracellular malondialdehyde （MDA） and myeloperoxidase （MPO） contents. The levels of caspase-3 and heat shock protein 70 （hsp70） mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. Results： The EDBD could protect the cardiomyocytes from H202 injury in a dose- dependent manner （3.75, 7.50, and 15.00 mg/mL）. The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. Conclusion： The EDBD has protection on cardiomyocytes injured by H202 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.

Danggui Buxue decoction ameliorates lipid metabolic defects involved in the initiation of diabetic atherosclerosis;identification of active compounds

OBJECTIVE: To determine the constituent compounds of Danggui buxue decoction(DBD) involved and the potential mechanisms mediating its effects, with specific reference to lipids playing a role in the initiation of diabetic atherosclerosis.METHODS: Liquid chromatography-tandem mass spectrometry was used to identify and quantify the absorbed bioactive compounds(ABCs) present in DBD. Goto-Kakizaki(GK) rats were randomly allocated to a diabetes atherosclerosis(DA) group, a DBD group, and an ABC group(10 per group), which were all high-fat diet-fed. The treated rats were administered DBD(4 g/kg) or ABCs(in amounts equal to those present in DBD) once daily for 28 d, and a control group of Wistar rats were administered vehicle. Body mass gain, fasting blood glucose, and homeostasis assessment of insulin resistance(HOMAIR) were measured. Serum triglyceride(TG), cholesterol(CHOL), high density lipoprotein-cholesterol(HDL-C), low-density lipoprotein-cholesterol(LDLC) and tumor necrosis factor-α(TNF-α) concentrations were determined. Hematoxylin and eosin staining and microscopy were used to characterize the abdominal aorta and the expression of lipogenic genes was quantified in this vessel.RESULTS: Seven ABCs were identified in rat serum:ferulic acid, formononetin, calycosin, astragaloside,caffeic acid, ligustilide, and butyphthalide. DBD significantly reduced HOMA-IR, the serum concentrations of TG, CHOL, and LDL-C, and the expression of the lipogenic genes monocyte chemotactic protein1, Fas, intercellular adhesion molecule 1, and Cd36 in aorta;and significantly increased the mRNA expression of Scd1 in aorta.CONCLUSION: DBD affects lipid metabolism in the early stage of atherosclerosis in diabetic GK rats,with the mechanism likely involving the regulation of lipid metabolic genes in vessels. The contribution of ABCs to the effect of DBD on lipid metabolism was 24%-101%.

Effects of Danggui Buxue Decoction (当归补血汤) on Lipid Peroxidation and MMP-2/9 Activities of Fibrotic Liver in Rats

Objective:To explore the mechanism of Danggui Buxue Decoction(当归补血汤,DBD) on the liver fibrosis related to hepatic lipid peroxidation and matrix metalloproteinases(MMP) -2/9 activities.Methods: The liver fibrosis in 28 rats was induced by an injection of carbon tetrachloride(CCl_4) and fed with high lipid and low protein diet for 6 weeks,the model rats were randomly divided into the model group and DBD treated group,14 in each group,and another 10 rats as the normal group were observed as well.Rats in the DB...

Efficacy of Auxiliary Therapy with Danggui Buxue Decoction No.1(当归补血汤1号) in Treating Patients of Non-small Cell Lung Cancer at Peri-operational Stage

Objective:To observe the therapeutic efficacy of Danggui Buxue Decoction No.1(当归补血汤1号,DB1) in treating patients of non-small cell lung cancer(NSCLC) at the peri-operational stage and its impact on the patients immune function.Methods:Eighty-two NSCLC patients were randomly assigned to two groups equally,the control group and the test group,they were given conventional treatment,while to the test group, DB1 were given additionally.The observation was conducted by testing the changes of T-lymphocyte subsets,...

Studies on blood enrichment and anti-tumor effects of combined Danggui Buxue Decoction,Fe and rhEPO based on colon cancer-related anemia model and gut microbiota modulation

Colon cancer-related anemia(CCRA)is mainly caused by systemic inflammation,intestinal bleeding,iron deficiency and chemotherapy-induced myelosuppression in colon cancer.However,the best therapeutic schedule and related mechanism on CCRA were still uncertain.Studies on blood enrichment and anti-tumor effects of combined Danggui Buxue Decoction(DBD),Fe and rhEPO based on CCRA and gut microbiota modulation were conducted in this paper.Here,CCRA model was successfully induced by subcutaneous inoculation of CT-26 and i.p.oxaliplatin,rhEPO+DBD high dosage+Fe(EDF)and rhEPO+DBD high dosage(ED)groups had the best blood enrichment effect.Attractively,EDF group also showed antitumor activity.The sequencing results of gut microbiota showed that compared to P group,the relative abundances of Lachnospiraceae and opportunistic pathogen(Odoribacter)in ED and EDF groups were decreased.Interestingly,EDF also decreased the relative abundances of cancer-related bacteria(Helicobacter,Lactococcus,Alloprevotella)and imbalance-inducing bacteria(Escherichia-Shigella and Parabacteroides)and increased the relative abundances of butyrate-producing bacteria(Ruminococcaceae_UCG-014),however,ED showed the opposite effects to EDF,this might be the reason of the smaller tumor volume in EDF group.Our findings proposed the best treatment combination of DBD,rhEPO and Fe in CCRA and provided theoretical basis and literature reference for CCRA-induced intestinal flora disorder and the regulatory mechanism of EDF.

Progress of Radix Astragali and Radix Angelicae Sinensis in the treatment of idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis(IPF)is a chronic,progressive,fibrotic interstitial lung disease.Current treatment options for IPF are limited.Radix Astragali(RA)and Radix Angelicae Sinensis(RAS),according to 5:1 ratio composed of Danggui Buxue decoction(DGBXD),which have played an essential role in the treatment of IPF.This article reviewed the experimental research,clinical research,and progress of RA and RAS(DGBXD)treating IPF to provide a deeper scientific basis for the future experimental research and clinical research.