Sodium Butyrate Can Inhibit Ferroptosis Through GPX4 And Improve Lipid Deposition Induced By Free Fatty Acids In Chicken Primary Hepatocytes

Fatty liver hemorrhagic syndrome(FLHS)is a common nutritional metabolic disease in laying hens during peak egg production,causing huge losses to the poultry industry.Previous studies in our group showed that the production of butyric acid,a metabolite of intestinal microorganisms,decreased during the formation of FLHS.However,whether dietary supplementation with sodium butyrate(Na B)can alleviate liver lipid deposition in FLHS laying hens has not been studied.Therefore,this paper aims to investigate the regulatory mechanism of Na B,the ferroptosis inhibitor Fer-1 and the activator RSL3 on the lipid metabolism and ferrosis of chicken primary hepatocytes induced by free fatty acids(FFAs),and to explore the regulatory mechanism of Na B on FFAs-induced hepatic liposis models.The results are as follows:1.FFAs induced increased the contents of TG,T-CHO,ALT and AST in chicken hepatocytes,the content of GSH-Px decreased,the content of Fe2+ and total iron increased,and the results of oil red staining showed that FFAs induced hepatocytes to present a large number of oil-red lipid droplets,the mitochondria became smaller under electron microscopy,the structure was blurred,the expression levels of GPX4,XCT,TFRC genes and proteins decreased,and the expression levels of NCOA4,FTH1 and DMT1 genes and proteins increased(P<0.05 or P<0.01).2.FFAs induced significant decrease in the contents of TG,ALT,AST,MDA,Fe2+under Na B treatment,oil red staining showed that lipid droplets in cells decreased,mitochondrial structure was clear under electron microscopy,mitochondrial crest increased,GPX4 and XCT expression levels increased,and NCOA4 and FTH1 expression levels decreased(P<0.05 or P<0.01).3.FFAs-induced hepatic lipid models under Fer-1 treatment,The contents of T-CHO,ALT,AST,Fe2+ and total iron were significantly reduced,the number of cell lipid droplets in the oil red result was reduced,and the expression levels of COX-2,FTH1 and NCOA4 decreased.The hepatolipid model was treated in combination with Fer-1 and Na B,compared with FFA+Na B group,the number of lipid droplets decreased,lipid deposition decreased,COX-2 and FTH1 expression levels decreased,and GPX4 expression levels increased(P<0.05 or P<0.01).4.FFAs-induced hepatic lipid models under RSL3 treatment,The contents of TG,ALT and AST increased significantly,the number of lipid droplets in the oil red results increased,and the expression levels of GPX4,NCOA4,FTH1 and XCT decreased.The hepatolipid model was treated in combination with RSL3 and Na B,compared with the FFA+Na B group,the number of lipid droplets increased,lipid deposition increased,the expression levels of GPX4 and XCT decreased,and the expression levels of COX-2 and ACSL4 increased(P<0.05 or P<0.01).Conclusion: FFAs induce fatty deposition in chicken hepatocytes;Na B can alleviate steatosis of chicken hepatocytes by inhibiting ferroptosis through GPX4.