视黄酸诱导H9c2细胞p38 MAPK转位的研究

Translocation of p38 MAPK induced by retinoic acid in H9c2 cells

目的了解H9c2心肌细胞中p38MAPK的分布及其在全反式视黄酸(atRA)诱导下的转位。方法对H9c2细胞进行饥饿加药,应用激光扫描共聚焦显微镜对细胞内的p38进行定位扫描。通过对p38荧光强度的核浆比进行分析,得出atRA与SB202190对H9c2细胞内p38核转位的影响。结果在未受刺激细胞内,p38较多分布在胞浆;经atRA分别刺激细胞5min,30min后,胞浆中p38的荧光强度均变弱,核浆比显示实验组与对照组间有显著性差异。SB202190能明显抑制atRA诱导的p38核转位(P<0.01),并且,由SB202190抑制组与对照组的p38核浆比能看出,SB202190降低了26.1%的p38核转位。结论atRA能诱导心肌细胞中p38的核转位,p38信号转导通路可能参与心脏的发育调控。

Objective To study the distribution of p38 MAPK and translocation of p38 induced by all-trans retinoic acid （atRA） in cardiomyocytes. Methods H9c2 cells were cultured in the normal growth medium containing 10% fetal bovine serum. Cells were serum-starved overnight followed by atRA （0.05μmol/L） treatment or SB202190 （10μmol/L） pretreatment. The localization of p38 under different conditions was detected by Confocal Laser Scanning Microscopy. Then the ratio of p38 localized in nuclei to cytoplasm was analyzed by a Leica Microsystem. Thus, the statistical analysis of these data showed the effects of atRA or SB202190 on p38 translocation. Results p38 was located more in the cytoplasm without stimulation, while in the cells with atRA treatment for 5 min and 30 min, p38 labeled with green fluorescence in the cytoplasm decreased. The p38 ratio of nuclei to cytoplasm showed that there was a significant difference between atRA groups and control. As the specific inhibitor of p38, SB202190 could obviously inhibited the nuclear translocation of p38 induced by atRA （P 〈 0.01 ）. And compared with control, SB202190 could make the nuclear translocation of p38 decrease by 26.1%. Conclusion atRA could induce the nuclear translocation of p38 in cardiomyocytes. And p38 pathway might participate in the regulation of heart development.