叶酸缺乏对鼠胚成纤维细胞MAPK通路及多潜能干细胞诱导效率的影响  

Influence of folic acid deficiency on mouse embryonic fibroblast MAPK pathway and induction efficiency of induced pluripotent stem cells

作　　者：胡文涛[1] 闫秋月[2] 方瑜[1] 邱占东[1] 张苏明[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院,武汉430030 [2]沧州市中心医院

出　　处：《内科急危重症杂志》2014年第1期51-53,共3页Journal of Critical Care In Internal Medicine

基　　金：国家自然科学基金资助项目(No:81271407)

摘　　要：目的:观察短暂性的叶酸缺乏对鼠胚成纤维细胞(MEF)MAPK通路及其诱导多潜能干(iPS)细胞诱导效率的影响。方法:将鼠胚成纤维细胞分为正常叶酸组、叶酸缺乏3 d组和叶酸缺乏6 d组,分别于叶酸缺乏3、6 d western blot法检测提取的上述3组鼠胚成纤维细胞中的磷酸化细胞外信号调节激酶(p-ERK)蛋白和总细胞外信号调节激酶(ERK)蛋白的变化,用经典四因子的逆转录病毒载体进行感染,并对获得iPS细胞进行鉴定。结果:叶酸缺乏3 d组p-ERK明显降低,感染后第12天出现具有胚胎干细胞形态的克隆,叶酸缺乏3 d组、6 d组产生的iPS细胞和正常叶酸组的诱导效率分别为(0.0477±0.0005)%、(0.0155±0.0019)%和(0.01±0.0005)%,所获的iPS细胞多能性基因和免疫荧光多能性标志鉴定为阳性。结论:叶酸缺乏3 d能明显压低p-ERK的表达,提高小鼠iPS细胞的诱导效率,且诱导的细胞多能性基因和免疫荧光多能性标志鉴定为阳性。Objective： To observe the influence of folic acid deficiency on the MAPK pathway of mouse embryonic fibroblast and induced efficiency of induced pluripotent stem（ iPS） cells. Methods： Mouse embryonic fibroblasts were divided into normal folic acid group,folic acid deficiency for 3 days and 6 days group. On the 3rd day and 6th day,western blot method was used to detect the changes of p-ERK and total ERK,three groups were infected with retroviruses expressing classic 4 factors. The obtained iPS cells were later identified. Results： Compared with that of the other groups,the p-ERK of folic acid deficiency for 3 days group was obviously inhibited,the embronic stem-like clones appeared on the 12th day after infection. Induced efficiency of the iPS cells for 3 days,6 days and control group was（ 0. 0477 ± 0. 0005） %,（ 0. 0155 ± 0. 0019） % and（ 0. 01 ± 0. 0005） % respectively. Pluripotency of the iPS cells obtained is positive. Conclusion： Folic acid deficiency for 3 days can obviously inhibit the expression of p-ERK and improve the induction efficiency of iPS cells. The iPS cells obtained are pluripotent,which may be related to inhibition of p-ERK expression.

关 键 词：叶酸缺乏 诱导多潜能干细胞 MAPK通路 逆转录病毒 

分 类 号：R73[医药卫生—肿瘤]