环格列酮和油酸对延边黄牛脂肪细胞分化的影响

Effects of Ciglitazone and Oleic Acid on Differentiation of Adipocytes of Yanbian Yellow Cattle

为探讨环格列酮和油酸对延边黄牛脂肪细胞分化的影响,试验采用胶原酶消化法在无菌条件下分离获得延边黄牛脂肪细胞,进行传代及诱导分化,观察细胞生长状态,绘制细胞生长曲线,通过油红O染色法及实时荧光定量RT-PCR方法研究环格列酮和油酸在延边黄牛脂肪细胞分化过程中的作用,对脂滴形成以及与脂肪细胞分化相关的主要标志基因PPARγ,SREBP1,C/EBPβ及SCD mRNA表达量的影响。结果表明:在不同分化处理144 h后,各处理组均可观察到有脂滴形成。其中,环格列酮单独处理时,显著提高了PPARγ、C/EBPβ和SCD基因mRNA的相对表达量(P<0.05);添加油酸后144 h,PPARγ基因的mRNA表达量与对照相比显著升高(P<0.05),SCD基因mRNA的表达量显著降低(P<0.05);同时添加环格列酮和油酸处理144 h后,PPARγ基因的mRNA表达量显著提高(P<0.05),但SREBP1和C/EBPβ及SCD基因的mRNA表达量无明显变化,说明在延边黄牛脂肪细胞的分化过程中,环格列酮和油酸对脂滴的形成及主要转录因子的表达量变化有着重要的调节作用。

To investigate the effects of ciglitazone and oleic acid on the differentiation of adipocytes of Yanbian Yellow Cattle,the adipocytes of Yanbian Yellow Cattle were isolated by collagenase digestion under aseptic conditions,and then subcultured and induced for differentiation.Oil red O staining and real time fluorescent quantitative RT-PCR method were used to plot cell growth and to check the effects of ciglitazone and oleic acid on adipocyte differentiation,lipid droplets and mRNA expression of PPARγ,SREBP1,C/EBPβand SCD.The results showed that after the treatment for 144 h,lipid droplets could be observed in all of the treatment groups,and the relative expression of PPARγ,C/EBPβand SCD genes was significantly increased when treated with ciglitazone alone,the expression of PPARγgene was significantly increased and the expression of SCD gene was significantly decreased after being treated with oleic acid.When the ciglitazone and oleic acid were added together after 144 h,the expression of PPARγgene was significantly increased,but there was no significant effect on the expression of SREBP1,C/EBPβand SCD genes.The results indicated that ciglitazone and oleic acid played an important role in the formation of lipid droplets and the expression of major transcription factors in the differentiation of adipocytes of Yanbian Yellow Cattle.