中国畜牧兽医2024，Vol.51Issue(7)：2766-2775,10.DOI:10.16431/j.cnki.1671-7236.2024.07.004
绿原酸对高游离脂肪酸处理犊牛肝细胞胆固醇和胆汁酸代谢的影响
Effects of Chlorogenic Acid on Cholesterol and Bile Acid Metabolism in Calf Hepatocytes Treated with High FFA
摘要
Abstract
[Objective]The objective of this study was to investigate the effects of chlorogenic acid(CGA)on cholesterol and bile acid metabolism in calf hepatocytes treated with high free fatty acid(FFA).[Method]The primary calve hepatocytes were separated by two-step collagenase perfusion method,and the cells were identified by immunofluorescence and divided into 4 groups.Cells in control group were cultured with RPMI-1640 medium containing 2％BSA.The cells in FFA group were cultured after adding 1.2 mmol/L FFA in RPMI-1640 medium containing 2％BSA,and CGA group were cultured after adding 20 μg/mL CGA in RPMI-1640 medium containing 2％BSA.Cells in CGA+FFA group were cultured after adding 1.2 mmol/L FFA and 20 µg/mL CGA in RPMI-1640 medium containing 2％BSA.Cells were collected after 12 h,and the contents of triglyceride(TAG)and total cholesterol(TC)were detected by the kit.Real-time quantitative PCR and Western blotting were used to detect the mRNA and proteins expression levels of cholesterol synthesis-related factors,including sterol regulatory element binding transcription factor 2(SREBF2)and 3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),and cholesterol effection-related factors,including acetyl-CoA acetyltransferase 2(ACAT2),ATP-binding box subfamily A member 1(ABCA1),ATP-binding box subfamily G member 5(ABCG5),and bile acid metabolism related factors,including cholesterol 7a-hydroxylase(CYP7A1),cholesterol 12a-hydroxylase(CYP8B1),cholesterol of 27 a-hydroxylase(CYP27A1),farnesol X receptor(FXR)and fibroblast growth factor receptor 4(FGFR4).[Result]After different treatments of calf hepatocytes,compared with control group,the TC content of calf hepatocytes in FFA group was extremely significantly reduced(P＜0.01),and the TAG content was extremely significantly increased(P＜0.01),and the mRNA expression levels of HMGCR,ABCA1,ABCG5,ABCG8,APOA1,ACAT1,NPC1L1,FXR and FGFR4 genes and the protein expression levels of SREBF2,ACAT2,ABCA1 and ABCG5 were extremely significant or significantly reduced(P＜0.01 or P＜0.05),the mRNA expression of CYP8B1 gene and the protein expression of CYP7A1 and CYP8B1 were extremely significant or significantly increased(P＜0.01 or P＜0.05).The TC content of calf hepatocytes in CGA group was extremely significantly increased(P＜0.01),the protein expression levels of SREBF2,ABCA1,CYP27A1,FXR and FGFR4 were extremely significantly increased(P＜0.01),the protein expression of CYP7A1 was extremely significantly reduced(P＜0.01).Compared with FFA group,the TC content of calf hepatocytes in CGA+FFA group was significantly increased(P＜0.05),and the TAG content was significantly reduced(P＜0.05),the mRNA expression levels of HMGCR,ACAT2,ABCA1,ABCG5 and APOA1 genes and the protein expression levels of SREBF2,ACAT2,ABCA1,ABCG5,FXR and FGFR4 were extremely significant or significantly increased(P＜0.01 or P＜0.05),the protein expression levels of CYP7A1 and CYP8B1 were extremely significantly reduced(P＜0.01).[Conclusion]CGA could participate in the regulation of intracellular cholesterol homeostasis in calf hepatocytes while activating FXR and FGFR4,which in turn alleviates high FFA-treated bile acid accumulation.关键词
绿原酸/游离脂肪酸(FFA)/犊牛原代肝细胞/胆固醇/胆汁酸Key words
chlorogenic acid/free fatty acid(FFA)/primary calve hepatocytes/cholesterol/bile acids分类
农业科技引用本文复制引用
田艳,杨威,刘佳晨,王爽,范文文,常雅奇,杨天娇,张泽鑫,张冰冰,徐闯..绿原酸对高游离脂肪酸处理犊牛肝细胞胆固醇和胆汁酸代谢的影响[J].中国畜牧兽医,2024,51(7):2766-2775,10.基金项目
黑龙江省自然基金优秀青年项目(YQ2022C029) （YQ2022C029）
