| Objective: The present research aimed to explore the effect and mechanism of astragalus membranaceus(AM)extract on hyperlipidemia,which will provide scientific basis for the development of new plant compounds to alleviate hyperlipidemia.Methods: The analysis methods of network pharmacology and bioinformatics were as follows:(1)the active components and targets of astragalus membranaceus were searched through traditional chinese medicine systems pharmacology database and analysis platform(TCMSP),comparative toxicogenomics database(CTD)and STITCH database;(2)The data set related to hyperlipidemia was retrieved from the gene expression omnibus(GEO)database,and the differentially expressed genes were screened by using the R language package "limma".Combined with the retrieval results of CTD,the targets related to hyperlipidemia were screened together;(3)The intersection of astragalus membranaceus and hyperlipidemia related targets was used to obtain candidate targets.The R language package "Cluster Profiler" is used to analyze the function enrichment of candidate targets to determine the main signal pathway;(4)The "astragalus membranaceus-component-target" network was further constructed by using Cytoscape software to screen core targets;(5)At the same time,the STRING database was used for protein-protein interaction analysis,and the GSEA software was used for gene set enrichment analysis to determine the key targets.In vivo animal experiment methods were as follows:(1)Adult C57BL/6J male mice were randomly divided into normal diet group(ND),high-fat diet group(HFD),and high-fat diet fed with astragalus membranaceus extract intervention group(HFD + AM).The food intake and body weight gain of mice was recorded;(2)After 16 weeks,blood samples were collected from eyeballs,and the levels of serumtotal cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)in serum were measured by chemical kit;(3)The contents of TC and TG in liver tissue,free fatty acid(FFA)and glycerol in white adipose tissue were detected;(4)Western blot was used to detect the expression of lipid metabolism related proteins in liver and white adipose tissue,including fatty acid synthase(FASN),fatty triglyceride lipase(ATGL)and carnitine palmitoyl transferase 1-α(CPT1-α);(5)Western blot was used to detect the expression of key targets in liver and white adipose tissue.Results: Through the analysis of network pharmacology and bioinformatics,the following results were obtained:(1)481 astragalus membranaceus related targets and 474 hyperlipidemia related targets were screened;(2)101 candidate targets were obtained from the intersection of astragalus membranaceus and hyperlipidemia related targets,which are mainly enriched in endocrine resistance,AGE-RAGE in diabetic complications and p53 signaling pathways.(3)The key targets of astragalus membranaceus in alleviating hyperlipidemia may be Akt serine/threonine kinase 1(AKT1),vascular endothelial growth factor A(VEGFA),cyclin D1(CCND1)and estrogen receptor 1(ESR1).These four key targets are closely related to adipogenesis,fatty acid metabolism and bile acid metabolism.The results of animal experiments showed that:(1)Without changing the food intake of mice(P>0.05),astragalus membranaceus extract greatly reduced weight gain in mice fed with high-fat diet(P<0.05);(2)astragalus membranaceus extract could significantly reduce the levels of serum TC,TG and LDL-C(P<0.05),and increase the level of serum HDL-C(P<0.05);(3)astragalus membranaceus extract significantly reduced the levels of TC and TG in liver(P<0.05)and the content of FFA in white adipose tissue(P<0.05).There was no significant difference in the intervention effect of astragalus membranaceus extract on the content of glycerol in white adipose tissue(P>0.05);(4)astragalus membranaceus extract could reduce the expression of adipogenesis related protein FASN in liver and white adipose tissue(P<0.05)and increase lipolytic protein ATGL and lipid β-Oxidation related protein CPT1-α(P<0.05);(5)After treated with astragalus membranaceus extract,the levels of AKT1 and CCND1 protein in liver and white adipose tissue of high-fat diet mice increased significantly(P<0.05),and the levels of VEGFA and ESR1 protein decreased significantly(P<0.05).Conclusion: astragalus membranaceus extract alleviated high-fat diet induced hyperlipidemia through regulating lipid metabolism,and AKT1,VEGFA,CCND1 and ESR1 might be the key targets. |
