Exploration of Effects of Buyang Huanwu Decoction(补阳还五汤)Containing Serum on High Glucose-Induced Vascular Endothelial Cell Injury Based on Network Pharmacology and Experimental Validation
Objective:To investigate the effects of Buyang Huanwu Decoction(补阳还五汤)containing serum on high glucose-induced vascular endothelial cell injury based on network pharmacology and experimental validation.Methods:Based on network pharmacology,the active chemical components and targets of Buyang Huanwu Decoction were obtained from TCMSP database,ChemSource and Swisstarget prediction databases,and diabetic vascular complications related targets were collected from GeneCards,OMIM and DrugBank databases.The PPI net-work of the key targets was constructed by using the String database.The GO and KEGG enrichment analysis were performed based on Metas-cape database.Molecular docking validation was performed using Autodock and Pymol software.A high glucose-induced HUVECS cell injury model was established,and 5％and 10％Buyang Huanwu Decoction-containing serum was given to the cells.CCK-8 assay was used to detect the cell proliferation of HUVECs,the TBA method was used to determine the MDA content,and the xanthine oxidase method was used to de-termine the SOD activity.Annexin V-FITC/PI double-labeling combined with Hoechst 33258 and JC-1 staining was used to detect apopto-sis.ELISA was used for the detection of IL-6 and TNF-α levels,and Western Blot was used to detect protein expressions of vascular endotheli-al growth factor(VEGF),p-Akt and Akt.Results:A total of 69 active ingredients and 306 potential targets were involved in the treatment of diabetic vascular complications with Buyang Huanwu Decoction.The main active ingredients involved quercetin,kaempferol,and β-sitosterol,and the key targets were vascular endothelial growth factor(VEGFA),cellular inflammatory factors,such as TNF and IL6,etc.The biological process involved the response to hormones,cellular response to nitrogen compounds,and cellular response to cytokines,etc.,and the cellular composition involved membrane rafts,membrane sides and receptor complexes,etc.The molecular function involved kinase activity and bind-ing,lipid binding,transcription factor binding,etc.The pathways of its possible mechanism of action may be related to lipid and atherosclero-sis,apoptosis and senescence,growth hormone synthesis and secretion,insulin resistance,AGE-RAGE signaling pathway in diabetes complica-tions,PI3K-Akt signaling pathway,etc.Based on the results of network pharmacology and experimental verification,compared with the model control group,the cell survival rate of the 18.5 g/kg Buyang Huanwu Decoction-containing serum 10％group was significantly increased(P＜0.01),and the MDA content of damaged endothelial cells was reduced(P＜0.05 or P＜0.01),while the SOD activity was increased(P＜0.01)in the 18.5 g/kg Buyang Huanwu Decoction-containing serum 5％and 10％groups.Compared with the model group,in the 18.5 g/kg Buyang Huanwu Decoction-containing serum 5％and 10％groups,cell apoptosis was significantly inhibited,and the contents of IL-6 and TNF-a were decreased in damaged endothelial cells.In addition,the protein expression of VEGF(P＜0.01),and p-AKT/AKT(P＜0.01)were down-regulated,and the intravascular environment under hyperglycemic state was improved.Conclusion:Buyang Huanwu Decoc-tion can improve the intravascular environment under hyperglycemic state in the treatment diabetic vascular complications,which may be relat-ed to its ability of protecting the endothelial cells in high glucose state,promoting cell proliferation,inhibiting apoptosis and inflammatory factor production,down-regulating the protein expression of VEGF and p-Akt/Akt,and regulating PI3k/Akt signaling pathway.
万方数据
维普
