Research On The Mechanism Of Biochanin A And Peptide CPe-Ⅲ Regulate The Lipid Metabolism

Abnormal lipid metabolism is one of the most important factors that lead to atherosclerosis and coronary heart disease.Eating the chickpea can effectively decrease the blood lipid level and reduce the accumulation of cholesterol.The nutritional components to reduce blood lipids are mainly concentrated on isoflavone(main biochanin A)and the protein / peptide.This paper systematically research the biological activity and structureactivity relationship of biochanin A and CPe-Ⅲ on the regulation of lipid metabolism by the chemical simulation system,cell culture,model mice experiment in vitro combined with molecular dynamics theory,which provide theoretical foundation and scientific basis for further research on natural active peptide,isoflavones for regulating lipid metabolism.Biochanin A was extracted from chickpea by the ultrasonic wave and purified by macroporous resin AB-8.The antioxidant activity was evaluated by the reducing power assay,DPPH,ABTS free radical scavenging ability.The results showed biochanin A had strong reducing power and free radical scavenging capacity with a dose effect relationship.Cell experiments showed biochanin A inhibited the proliferation of tumor cells.Biochanin A can accelerate the cholesterol efflux in RAW264.7 macrophages,reduce the accumulation of cholesterol,and inhibit the forming of foam cell.To establish model of mice with hyperlipidemia,with simvastatin as positive control,the mice were determined the weight,fecal fat,total cholesterol,triglyceride,high density lipoprotein,low density lipoprotein,hepatic lipase activity and organ index in liver,heart,spleen,kidney and epididymal fat and observed the liver by HE staining to systematically research the biochanin A and CPe-Ⅲ on the regulation of lipid metabolism.The results showed that compared to the high-fat-diet group,the triglyceride and total cholesterol of each group were lower;high density lipoprotein content increased;the relative enzyme such as SOD,LPL,HL and fecal fat increased;the liver pathological changes were also improved accordingly.Biochanin A and peptide CPe-Ⅲ can effectively improve the abnormal lipid metabolism of the KM mice with hyperlipidemia.Based on the analysis of the binding site of biochanin A and CPe-Ⅲ with the CETP targets,molecular docking was analyzed by using Autodock,ZDOCK,FiberDock software,and molecular dynamics simulation was analayzed the interaction by Amber.The results showed that the active region of biochanin A and receptor CETP mainly distributed in the two hydrophobic regions,which can be effectively combimed and interacted with the ligand lipophilic side chain or group;CPe-Ⅲ combined with CETP in the narrowest place in the channel,which hindered the reverse lipid transport,increased HDL and facilitated the lipid metabolism.