FCCP对P19细胞向心肌细胞诱导分化的影响

目的:探讨羰基-氰-对-三氟甲氧基本腙(carbonyl cyanide p-trifluoromethoxyphenylhydrazone,FCCP)对P19细胞向心肌细胞诱导分化的影响?方法:在P19细胞向心肌细胞诱导分化的过程中加入5 umol/L的FCCP,同时设立阴性对照组,显微镜观察分化过程中不同时间点P19细胞形态的变化,并采用 real-time PCR和蛋白质印迹法分别检测GATA4?NKX2.5及cTnT在分化过程中的mRNA和蛋白表达水平?结果:FCCP处理组P19细胞形成的胚胎样小体较对照组体积小,结构较松散,形态不均一?在整个分化过程中,FCCP处理组GATA4 ?NKX2.5及cTnT基因均有表达,但整个动态表达过程较对照组延迟;Western blot结果显示,FCCP处理组GATA4?NKX2.5蛋白在第5?7?9天的表达量均较对照组低;在第11天,两组间GATA4蛋白的表达量差异无统计学意义;而NKX2.5在FCCP处理组的表达量仍低于对照组?FCCP处理组和对照组cTnT蛋白在分化第5?7天的表达量差异无统计学意义,而在第9?11天cTnT蛋白表达量FCCP组均较对照组低?结论:FCCP可抑制P19细胞向心肌细胞诱导分化的进程?

Effect of FCCP on differentiation of P19 cell to cardiomyocytes

Objective:To investigate the effect of FCCP on differentiation of P19 cells to cardiomyocytes. Methods:P19 cells were treated with 5 umol/L FCCP and untreated cells were used as controls,the morphologic changes of P19 cells at indicated time during the differentiation were observed and photographed under an inverted microscope. The expressions of GATA4,NKX2.5 and cTnT were detected using Real-time PCR and Western blot in the differentiated process. Results:Compared to the control group,the embryoid bodies formed in the FCCP treated group were smaller and were loosely structured. The expressions of GATA4,NKX2.5 and cTnT were observed in the two groups,while FCCP treated cells present with a slower dynamic changes during the differentiation. Western blotting results showed that the protein expressions of GATA4,NKX2.5 in the FCCP treated group on day 5,7 and 9 were lower than that of the control group,respectively. For the expression of GATA4 protein on day 11,there was no significant difference between the two groups,while the expression of NKX2.5 protein in the FCCP treated group was lower compared with the control group. The protein expressions of cTnT on day 5,7 showed no significant difference within the two groups,and with further differentiation,the expression of cTnT protein in the FCCP treated group increased but was still lower than the control group on day 9,11,respectively. Conclusion:FCCP inhibited the differentiation of P19 cells to cardiac myocytes.