The Investigation On Kelationsinip Between AMPK Signal Pathway In Lipid Metabolism In Primary Hepatocytes From Laying Hens

To explore the role of AMPK signaling pathway in lipid metabolism in primary liver cells of laying hens.The high-activity chicken primary hepatocytes were obtained by modified in situ two-step perfusion method,and then the liver cells were treated with fatty acid(oleic acid:palmitic acid 2:1)to establish a steatosis model by referring to the non-alcoholic fatty liver in vitro modeling method.,divided into normal liver cell group(Control group),normal liver cell+AICAR group(Con+AICAR group),normal liver cell+Compoud C group(Control+Comp C group),steatosis liver cell group(FFA group),The steatosis hepatocytes+AICAR group(FFA+AICAR group),steatosis liver cells+compound C group(FFA+comp C)six groups.Normal hepatocytes and steatosis hepatocytes were treated with AMPK activator AICAR and inhibitor Compoud C,respectively,and blood lipids in all groups of hepatocytes were determined(triglyceride TG,total cholesterol TC,low density lipoprotein LDL-ch).,high-density lipoprotein HDL-ch),and mRNA expression levels of AMPK signaling pathway-related genes and lipid-cholesterol-related genes(AMPK?1,LKB1,ACC;FAS,HNF4a,HMGR,PPARa;CPT1,GAPT).The results are as follows:1.Improve the liver culture system of the original laying hens.The nucleus of the cultured primary chicken liver cells is clear,and the cell differentiation is completed after 24 hours of culture,and the cell viability index is high.The successful establishment of FFA(oleic acid/palmitic acid)-induced fat degeneration model of the laying hens showed that the lipid droplet area increased in a period of time,and the amount of lipid droplets increased significantly at 24 h(P<0.05).3.0.5 mM AICAR can activate AMPK at 1.5 h,and the activity is the highest;while 20 ?M Compoud C inhibits AMPK at 24 h,and the effect is most obvious.Compared with the Hepatocyte group in the Control group,the levels of TC,TG,LDL-ch in the FFA group increased significantly,and the HDL-ch content decreased significantly(P<0.05).The Control+AICAR group and the FFA+AICAR group and the Control group,respectively.Compared with FFA group,TG,TC content decreased significantly(P<0.05),and HDL-ch content increased(P<0.01);Control+Comp C group and FFA+ComC group compared with Control group and FFA group,TG and The TC content increased,but the difference was not significant(P>0.05),the LDL-ch content also increased but the difference was not significant,and the difference of HDL-ch content decreased significantly(P<0.01).Compared with normal hepatocytes,the expression of HMGR,ACC,FAS,GAPT,CPT1,HNF4a and PPARa mRNA in fatty degeneration liver cells increased significantly(P<0.05).The mRNA expression levels of AMPKal,LKB1 and CPT1 were significantly decreased(P<0.05).The expression levels of LKB1,AMPK?1,CPT1,HMGR,FAS and PPARa were compared between Control+AICAR group and FFA+AICAR group compared with Control group and FFA group.Significantly increased(P<0.05),GAPT,HNF4a mRNA expression decreased significantly(P<0.05),Control+Comp C group and FFA+ComC group compared with Control group and FFA group,LKB1,AMPKal,CPT1,HMGR,FAS The expression of PPARa mRNA was significantly decreased(P<0.05),and the expression of GAPT and HNF4a mRNA was significantly increased(P<0.01).The difference of HMGRa mRNA was not significant(P>0.05),but it decreased.Conclusion:Oleic acid/palmitic acid can cause simple steatosis in adult liver hens;and the expression of cholesterol synthesis gene is altered by activating AMPK signaling pathway;AMPK exogenous activators and inhibitors can be regulated The expression activity of AMPK in the liver cells of laying hens regulates the fat metabolism pathway of cholesterol synthesis.