ADIG诱导牛成肌细胞转分化及相关基因的表达分析

ADIG Inducing the Trans-differentiation of Bovine Myoblasts and Its Related Gene Expression

旨在通过在牛成肌细胞中过表达ADIG基因,诱导成肌细胞向成脂转分化,探讨该牛脂肪分化转录因子ADIG基因在脂类代谢过程中的功能,为阐明牛脂肪细胞分化转录因子ADIG基因在牛脂肪组织生长发育过程中发挥的作用提供证据。本研究用AD-ADIG腺病毒原液诱导转分化初生荷斯坦牛的成肌细胞,并在诱导分化后第2、4、6、8、10、12、14天进行油红O染色观察诱导成脂情况,同时收集细胞提取总RNA,利用实时荧光定量PCR分别检测ADIG、PPARγ、C/EBPα、SREBPF1、FABP4、FAS、MyoD基因的表达量变化。实时荧光定量PCR检测结果表明,ADIG基因在第2、4、6、8、10、12、14天均有高水平表达,且随着诱导天数增加脂滴明显增多。成脂相关的基因PPARγ、SREBPF1和FAS在后期AD-ADIG组中表达量均高于空白组,成肌相关的基因MyoD相对表达量在初期就显著下降,且AD-ADIG组明显低于对照组。综合脂滴染色和相关基因表达量方面的研究结果,说明ADIG基因在诱导牛成肌细胞分化向成脂转分化过程中发挥着重要作用,而且牛成肌细胞有向脂肪细胞分化的倾向。

The aim of this study was to provide evidence about the role of ADIGgene in bovine adipose tissue during growth and development,and to lay a theoretical basis for further study of the molecular mechanisms of bovine fat metabolism and meat quality improvement through inducing bovine myoblasts trans-differentiating into fat with overexpressing ADIG gene in bovine myoblasts.We induced bovine myoblasts trans-differentiating with AD-ADIG recombinant adenovirus,observed the lipid droplets after Oil Red O staining,collected cells to extract RNA at the 0th,2nd,4th,6th,8th,10th,12th,14 th day,respectively,and detected relative expressions of fat-related genes ADIG,PPARγ,C/EBPα,SREBPF1,FABP4,FAS as well as the muscle-related gene MyoDby real-time quantitative PCR.The real-time quantitative PCR results showed that during the induced trans-differentiation process,ADIGgene was expressed at high levels at the 2nd,4th,6th,8th,10 th,12th,14 th day,and lipid droplets increased significantly.The relative expression of the fat-related genes PPARγ,SREBPF1 and FASat late stages were obviously higher in AD-ADIG group than that in other groups,and the relative expression of muscle-related gene MyoD de-creased at early stages,and was obviously lower in AD-ADIG group than that in other groups.The result indicate that ADIGgene play important roles in bovine myoblasts induced into fat and bovine myoblast has the potential of differentiating to adipocyte according to the results of Oil Red O staining and related gene relative expression.