PPARγ激动因子与胰岛素互作影响延边黄牛前脂肪细胞分化  

Interaction between PPARγagonist and insulin affects the differentiation of preadipocytes in Yanbian yellow cattle

作　　者：郭盼盼 闫妍 张军芳 金鑫[1] 李香子[1] 严昌国[1] 尹云厚[2] GUO Panpan;YAN Yan;ZHANG Junfang;JIN Xin;LI Xiangzi;YAN Changguo;YIN Yunhou(College of Agriculture,Yanbian University,Yanji,Jilin 133000,China;Guizhou Minzu University,Guiyang 550025,China)

机构地区：[1]延边大学农学院,吉林延吉133000 [2]贵州民族大学,贵州贵阳550025

出　　处：《中国兽医学报》2021年第6期1179-1186,共8页Chinese Journal of Veterinary Science

基　　金：贵州省科技计划资助项目(黔科合基础[2018]1073)。

摘　　要：本试验旨在研究胰岛素和PPARγ激动因子对延边黄牛前脂肪细胞脂质代谢的影响。利用胶原酶消化法在无菌条件下分离获得延边黄牛原代脂肪细胞并进行体外培养,试验分为对照组(10mg/L胰岛素)、IC组(10mg/L胰岛素+10mg/L环格列酮),IO组(10mg/L胰岛素+100μmol/L油酸)和ICO组(10mg/L胰岛素+10mg/L环格列酮+100μmol/L油酸),通过油红O染色法、甘油三酯含量测定及实时荧光定量RT-PCR等方法检测诱导分化处理96h后,脂滴生成情况以及对脂肪细胞分化相关的主要标志基因PPARγ、SCD、LPL、SREBP1和C/EBPβmRNA表达的影响。油红O染色和甘油三酯测定结果显示,各试验组均可促进脂滴的形成和甘油三酯的积累,且相比于对照组和IC组,添加胰岛素和油酸的处理可更有效地促进延边黄牛前脂肪细胞脂滴的形成和甘油三酯含量的积累。实时荧光定量PCR结果显示,胰岛素与环格列酮和油酸共处理可提高PPARγ、SCD、LPL和C/EBPβ的表达量,胰岛素与油酸共处理在提高PPARγ、C/EBPβ和SREBP1基因mRNA表达量的同时,降低了SCD和LPL基因mRNA的表达,胰岛素与环格列酮共处理均显著提高了各基因mRNA的表达量(P<0.05)。试验结果为研究胰岛素、环格列酮和油酸三者互作效果在调节脂肪生成中的功能和分子机制提供了依据。This study investigated the effect of insulin and PPARγagonists on lipid metabolism in preadipocytes of Yanbian yellow cattle.The primary fat cells of Yanbian yellow cattle were isolated by collagenase digestion and cultured in vitro,and 10mg/L insulin+10mg/L cycloglitazone(IC group),10mg/L insulin+100μmol/L oleic acid(IO group),10mg/L insulin+10mg/L cycloglitazone+100μmol/L oleic acid(ICO group)were added to the differentiation medium and cells were cultured for 96h,the lipid droplets and the expression of PPARγ,SCD,LPL,SREBP and C/EBPβmRNA were detected by oil red O staining,triglyceride content determination and real-time quantitative PCR.The results of oil red O staining and triglyceride determination showed that,compared with the control group and IC group,the treatment of insulin and oleic acid could effectively promote the formation of lipid droplets and the accumulation of triglyceride content in preadipocytes.The results of real-time quantitative PCR showed that the expression of PPARγ,SCD,LPL and C/EBPβincreased when cells were treated with insulin and cycloglitazone and oleic acid.The co-treatment of insulin and oleic acid increased the mRNA expression of PPARγ,C/EBPβand SREBP1,but decreased the mRNA expression of SCD and LPL.The co-treatment of insulin and cycloglitazone significantly increased the mRNA expression of each gene(P<0.05).This study provides a basis for the study of the function and molecular mechanism of insulin,cycloglitazone and oleic acid in regulating adipogenesis.

关 键 词：延边黄牛 前脂肪细胞 胰岛素 环格列酮 油酸 

分 类 号：S823.2[农业科学—畜牧学]