Study On The Mechanism Of Fatty Acid Metabolism By Genistein

Abnormal fatty acid metabolism is one of the important factors of nonalcoholic fatty liver disease(Nonalcoholic fatty liver disease,NAFLD).As a kind of chronic disease,NAFLD is often accompanied by hepatocyte degeneration,triglyceride deposition,blocked-fatty acid oxidation,excessive fatty acid synthesis and inflammatory reaction.Genistein(Gen)is the main active substance of soybean isoflavones,which has a variety of biological functions and is widely involved in the regulation of human lipid metabolism,fatty acid metabolism and other physiological activities.Experimental results show that gen can activate PPARs and carbohydrate response element binding protein(ChREBP).It plays a vital role in regulating the expression of key target genes,the content of key enzymes,the level of related cytokines.Objective:Studying the effects of Gen on the expression of PPARs,Carbohydrate reaction element binding protein(ChREBP),Carnitine acetyltransferase 1(CPT-1),acetyl-CoA carboxylase(ACC)and the regulatory mechanism CPT-1,ACC and Malonyl-CoA(M-CoA)via fatty acid metabolic signaling pathway.Investigating the effects of gen on the content of triglyceride(TG),free fatty acid(FFA),and the level of inflammatory cytokines;It is helpful to guide the rational administration of drugs in clinic,to reduce or prevent the side effects of drugs,to provide scientific basis for improving the abnormal metabolism of fatty acids.The research on the mitigation and prevention of liver metabolic diseases such as NAFLD provides experimental basis and has important significance in many aspects such as the prevention of NAFLD disease and the development of new drugs.Methods:Cells(Hep G2,Hep 3b,L-02,3T3-L1)are cultured in vitro and induced by the dose of 1mM OA to generate fat deposition and to establish NAFLD cell model.The experiment consists of three parts:firstly,cells be divided into blank group,model group,Gen groups,PPARs agonist groups.According to the indexes measured before and after adipose,the regulatory effect of gen was analyzed by synthesizing comprehensive analysis of various factors after measuring the content of FAA and TG;Second,PPARs,ChREBP,CPT1 and ACC mRNA expressions are determined by real-time fluorescence quantitative PCR;the content of malonyl coenzyme A is detected by Double antibody sandwich method(ELISA).To investigate the effects of gen,PPARs agonists F and R on PPARs,ChREBP,CPT1,ACC and M-CoA.To research Regulatory effect of gen on metabolic axis composed of CPT1,ACC and M-Co A;Finally,Oleic acid induced 3T3-L1 on the basis of adipogenic cell model,cells are treated with Gen,PPARs activators F and R,after that the levels of inflammatory factors(TNF-?,Leptin)are mensurated by ELISA.Results:(1)According to the levels of ALT and AST and cell viability at different concentrations of oleic acid,1mM OA is suitable for the establishment of cell NAFLD model in vitro.Gen cam effectively regulate the contents of triglycerides and free fatty acids which were significantly dependent on Gen.The higher Gen concentration,the lower the TG and FAA level.(2)Gen can up-regulate the expression of PPARs and its target gene CPT-1 and down-regulate ChREBP and its target gene ACC.Gen can significantly reduce the level of malonyl CoA in NAFLD model.(3)Levels of TNF-a and Leptin are related to the concentration of Gen.The higher the concentration of Gen,the less the concentration of TNF-a and Leptin.Conclusion:1 mM OA is suitable for the establishment of cell NAFLD model in vitro.Gen can affect the mRNA expression of ChREBP,CPT-1,ACC,the content of malonyl coenzyme A,and the level of inflammatory cytokines via PPARs signaling pathway.and it exists a negative correlation between the expression of CPT-1 and the malonyl coenzyme A via PPARs signaling pathway.Gen can Promote fatty acid oxidation,inhibit fatty acid synthesis,regulate inflammatory reaction.Accordingly,Gen reduces the accumulation of free fatty acids and triglycerides in cells.The experiment has important physiological and clinical significance to develop low toxicity and no side effect drugs of NAFLD and to prevent and alleviate NAFLD.