抑制NHE1活性对干细胞向心肌分化的影响

Effect of NHE1 on Stem Cell Differentiation into Cardiomyocytes

摘要:

Na+/H+交换蛋白1(NHE1)在心肌细胞发育过程中发挥重要的调节功能。为深入探索NHE1活性对干细胞向心肌分化过程中产生的影响, 本研究采用二甲基亚砜(DMSO)诱导P19干细胞向心肌细胞分化, 同时在培养液中添加NHE1抑制剂EMD87580, 对诱导后形成的类胚体进行检测。通过细胞形态观察、免疫组织化学染色及检测心肌特异表达基因等方法证明, 经诱导形成的类胚体贴壁生长后, 会向心肌细胞分化并出现跳动细胞团。而经过抑制剂处理的P19干细胞尽管能够形成类胚体且贴壁培养后细胞仍具有增殖活力, 细胞团周边也较整齐, 但未出现向心肌细胞分化的现象。这一结果表明, 抑制NHE1的活性, 能够影响P19干细胞向心肌细胞的分化作用。

Abstract:

Sodium/proton exchanger 1 (NHE1) plays an important role in the cardiomyocyte development. To study the effect of NHE1 activity in stem cells differentiation into cardiomyocytes, we treated P19 stem cells with dimethyl sulfoxide (DMSO) to initiate cardiomyocyte differentiation. In separate experiments, P19 cells were incubated with NHE1 specific inhibitor EMD87580 during the DMSO induction. The formed embryoid bodies (EBs) were detected with cell morphology detection, immunohistochemisty staining and RT-PCR analysis of expression of cardio-specific gene markers. Results showed that P19 cells were able to differentiate into cardiomyocytes and form the beating cell clusters. However, when cells treated with NHE1 inhibitor EMD87580, they could still form the EBs and proliferate when cell clusters adhered on the culture plate, but cells were unable to differentiate. This observation indicates that inhibition of NHE1 activity affected P19 stem cells differentiating into cardiomyocytes.