摘要
为研究FoxO1与骨骼肌纤维类型之间的关系,本试验以大白猪为实验材料,利用RT-PCR和Western印迹技术,检测了FoxO1与肌纤维类型标志基因MyHCⅠ、MyHCⅡa、MyHCⅡb和MyHCⅡx在特定骨骼肌中的表达规律,以及调控肌纤维类型关键基因Mef2c和NFAT的表达,并用Wortmannin处理原代培养的猪骨骼肌成肌细胞,检测了FoxO1与肌纤维类型相关基因的表达.结果显示,FoxO1在不同骨骼肌类型中mRNA表达差异不显著(P>0.05),其蛋白表达与MyHC各亚型显著相关.Wortmannin处理结果显示,在处理的第3和5d,FoxO1蛋白与MyHCⅡb,MyHCⅡx和NFAT表达显著正相关,而与MyHCⅠ,MyHCⅡa和Mef2c表达显著负相关.结果表明,FoxO1通过抑制MyHCⅠ的表达调控肌纤维类型.
To investigate the role of FoxO1 on the determination of Yorkshire porcine skeletal muscle fiber types,the FoxO1 expression was assayed by RT-PCR and Western blot.The marker or involved genes for skeletal muscle fiber type regulations,such as MyHC Ⅰ,MyHCⅡa,MyHCⅡb and MyHCⅡ x,NFAT and Mef2c were also characterized.Wortmannin was used to enhance the FoxO1 activity in skeletal muscle myoblast to further explore the role of FoxO1 on muscle fiber type regulation.No significant difference（P0.05） of FoxO1 mRNA levels was observed in various types of skeletal muscles,however,the FoxO1 protein level correlated with the expressions of MyHC isoforms.With Wortmannin treatments,the FoxO1 protein expression was positively correlated with MyHCⅡb and MyHC Ⅱx in primary myoblast collected on day 3 and day 5.A negative correlation of FoxO1 with MyHCⅠand MyHCⅡa was observed in contrast with the Mef2c and NFAT expressions.We postulated that Mef2c and NFAT might mediate the FoxO1 activity to regulate muscle fiber types.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2010年第3期283-289,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家重点基础研究发展计划(973计划
No.2004CB117506)资助~~
