The Mechanism Of Ferulic Acid,one Of Major Chemical Ingredients Of DXR Ⅲ On Atherosclerosis Based On Gut Microbiota And Lipid Metabolism

Background and objective:Cardiovascular and cerebrovascular diseases,which mostly develop from atherosclerosis,have become the leading cause of death worldwide.The disorder of Lipid metabolism has been considered the role pathological mechanism involved in the pathogenesis of atherosclerosis.Recently,a growing body of evidence has revealed that the gut microbiota is closely associated with the development of atherosclerosis.Dingxin Recipe Ⅲ(DXR Ⅲ)is a Chinese herbal compound,which be used for AS treatment for 20 years.According previous studies,the chemical composition such as Geniposide,Danshensu,Ferulic acid(FA)and Berberine are including in DXR Ⅲ.However,the mechanisms of DXR Ⅲ and FA in AS treatment are not clear.Based on the gut microbiota and lipid metabolism,we aimed to investigate the effect and mechanism of DXR Ⅲ and FA in AS treatment.Methods:1.Male SD rats were fed a high-fat diet(HFD)for 6 weeks and then DXR Ⅲwere administered by gavage for 4 weeks.The effects of DXR Ⅲ on serum lipids and liver lipid deposition were investigated.2.The chemical composition of DXR Ⅲwas analyzed by UHPLC-QE-MS.3.AS model was established by ApoE-/-mice which fed on HFD for 12 weeks,then ApoE-/-mice were gavaged with FA for another 12 weeks.The changes of atherosclerotic plaque and liver pathologies were observed using oil-red O,H&E and Masson staining.Serum levels of lipid and Alanine aminotransferase(ALT),Aspartate aminotransferase(AST)of the mice were examined.The protein and mRNA levels of lipid metabolism related genes were determined with qPCR,western blotting.Mice feces were collected to detect gut microbiota by 16S rRNA sequencing,and fecal metabolites by 1H NMR spectroscopy.Spearman correlation analysis was used to further examine the connection between atherosclerosis,gut microbiota and fecal metabolites.Results:1.DXR Ⅲ treatment significantly down-regulated serum lipid levels and improved mitigating hepatocyte steatosis in hyperlipidemia rats.2.The chemical composition of DXR Ⅲ including Geniposide,Danshensu,Ginsenoside,FA,Chlorogenic acid and so on.3.In ApoE-/-mice,FA significantly reduced the atherosclerosis plaque area.Meanwhile,FA regulated the serum lipid levels in AS mice.Liver injury and hepatocyte steatosis induced by HFD were also mitigated by FA.Furthermore,FA improved lipid metabolism possibly through up-regulation of AMPKa phosphorylation and down-regulation of SREBP1 and ACC1 expression(P<0.05).We also found that FA induced marked structural changes in the gut microbiota such as reducing reduced the relative abundance of Fimicutes,Erysipelotrichaceae and Ileibacterium(P<0.05).Meanwhile,FA up-regulated cholate and down-regulated acetate,alanine in fecal metabolites(P<0.05).The regulation of FA in gut microbiota and fecal metabolites were positively correlated with serum lipid levels in atherosclerosis mice.Conclusion:1.DXR Ⅲ treatment significantly alleviates hyperlipidemia in rats.2.The chemical composition of DXR III includes Geniposide,Danshensu,Ginsenoside,FA,Chlorogenic acid and so on.3.FA significantly protects against atherosclerosis probably through modulation of gut microbiota and lipid metabolism,likely involving activation of the AMPKα/SREBP 1/ACC 1 pathway.