微小RNA对心肌细胞C反应蛋白功能水平的调节

目的探讨微小RNA(microRNA,miRNA)基因调节机制对冠心病标志性分子C反应蛋白(CRP)的调节。方法以大鼠心肌细胞H9c2(2-1)为实验对象,在培养中进行miR312的增强和抑制处理:加入Pre-miR-132作为miR-132增强剂,Pre-miR miRNA前体分子阴性对照1号作为增强处理的对照;加入Anti-miR-132作为miRNA-132抑制剂,Anti-miR miRNA抑制剂阴性对照1号作为抑制处理的对照。用iFect转染后培养48 h,收集细胞进行CRP的蛋白免疫印迹杂交。结果大鼠心肌细胞株H9c2(2-1)在增强剂Pre-miR-132处理48 h后,CRP蛋白水平明显下降(t-检验,P=0.013 21);与此同时,抑制剂Anti-miR-132处理48 h后,CRP蛋白水平出现了明显上升(t-检验,P=0.012 40)。结论微小RNA miR-132对大鼠心肌细胞CRP功能水平具有显著的调节作用,二者呈负相关关系。

Regulation of a MicroRNA to the Functional Levels of Myocardial C-reactive Protein

Objective To investigate the regulation of microRNA（miRNA） to C-reactive protein,which is a marker molecule in coronary heart diseases.Methods Rat myocardial cell line H9c2（2-1）in culture was subjected to miR-132 enhancing and inhibiting treatments.Pre-miR-132 was applied to boost the functional levels of miR-132 and controlled with the Pre-miR miRNA precursor molecule negative control 1;Anti-miR-132 was applied to blunt the functional levels of miR-132 and controlled with the Anti-miR miRNA inhibitor negative control 1.The precursor and inhibitor were introduced to H9c2（2-1）cells using iFect mediated transfection.The treatments lasted for 48 hours,cells were then harvested for CRP immunoblotting analysis.Results 48 hours treatment of Pre-miR-132 significantly decreased the CRP levels in H9c2（2-1）cells（t-test,P=0.013 21） while 48 hours treatment of Anti-miR-132 significantly increased the CRP levels in H9c2（2-1）cells（t-test,P=0.012 40）.Conclusion microRNA miR-132 possesses a significant regulative role to rat myocardial CRP levels,and shows a pattern of negative correlation.