小鼠缺血性脑卒中后microRNA-128的表达及其对p38α MAPK调控作用的研究-东南大学学报医学版

小鼠缺血性脑卒中后microRNA-128的表达及其对p38α MAPK调控作用的研究

单位：西安交通大学医学部第三附属医院神经外科, 陕西西安 710061

分类号：R-33;R743.31

出版年·卷·期（页码）：2017·36·第二期（182-187）

摘要：

目的：探讨p38α蛋白在脑缺血后2 h降低的原因以及p38α蛋白水平表达是否受调于microRNA-128。方法：首先通过qPCR和Western blotting来检测p38α mRNA和蛋白的表达水平，其次通过生物信息学软件预测出p38α的表达可能受调于microRNA-128，最后进行细胞水平验证，使用携带有microRNA-128序列质粒的慢病毒载体、携带有microRNA-128反义序列质粒的慢病毒载体以及相应空质粒的慢病毒载体转染SH-SY5Y细胞，观察p38α蛋白的表达情况。结果：小鼠脑缺血后2 h p38α mRNA水平无降低，但其蛋白水平却明显下降，microRNA-128的表达水平明显升高；转染实验发现，转染microRNA-128序列质粒的慢病毒载体的细胞p38α蛋白水平明显降低，转染microRNA-128反义序列质粒的慢病毒载体的细胞p38α蛋白水平明显升高。结论：脑缺血后p38α蛋白的表达水平下降并不是因为p38α mRNA水平的降低，microRNA-128可以调控p38α蛋白的表达，可能是通过结合p38α的mRNA从而抑制其蛋白的翻译表达。

Objective: To investigate the reason of down-regulation of p38α protein 2 h after cerebral ischemia, whether the expression of p38α protein is regulated by mcroRNA-128. Methods: First, the expression levels of p38α mRNA and protein were detected by qPCR and Western blotting respectively, and than some predictions were done by using bioinformatics software, and we found that microRNA-128 may target p38α. Finally, the cellular level verification was conducted by using lentiviral vector carrying microRNA-128 sequence plasmid, microRNA-128 antisense sequence plasmid and empty plasmid to transfect SH-SY5Y cells. Results: p38α mRNA levels were not reduced 2 h after ischemia, while its protein level was significantly decreased, and microRNA-128 expression levels significantly increased; transfection experiments found that p38α protein levels which cells were transfected with lentiviral vector carrying microRNA-128 sequence plasmid were significantly lower, but p38α protein levels were significantly increased by transfecting lentiviral vector carrying microRNA-128 antisense sequence plasmid. Conclusion: The decline of p38α protein expression after cerebral ischemia is not due to the reduced level of p38α mRNA, microRNA-128 can regulate the expression of p38α protein, which may inhibit its protein expression by binding p38α mRNA.

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