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

发布时间：2019-01-05 15:39

【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p38αmRNA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 p38αmRNA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p38αmRNA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。
[Abstract]:Objective: to investigate the causes of the decrease of p38 伪 protein at 2 h after cerebral ischemia and whether the expression of p38 伪 protein is regulated in microRNA-128.. Methods: firstly, the expression of p38 伪 mRNA and protein was detected by qPCR and Western blotting, and then the expression of p38 伪 was predicted by bioinformatics software. The lentivirus vector carrying microRNA-128 sequence plasmid, the lentivirus vector carrying microRNA-128 antisense sequence plasmid and the corresponding empty plasmid lentivirus vector were used to transfect SH-SY5Y cells to observe the expression of p38 伪 protein. Results: at 2 h after cerebral ischemia, the level of p38 伪 mRNA did not decrease, but the protein level decreased significantly, and the expression of microRNA-128 increased significantly. The p38 伪 protein level of lentivirus vector transfected with microRNA-128 sequence plasmid was significantly lower than that of lentivirus vector transfected with microRNA-128 antisense sequence plasmid. Conclusion: the decrease of p38 伪 protein expression after cerebral ischemia is not due to the decrease of p38 伪 mRNA level. The expression of p38 伪 protein can be regulated by microRNA-128, which may inhibit the translation of p38 伪 protein by binding p38 伪 mRNA.
【作者单位】：西安交通大学医学部第三附属医院神经外科;
【基金】：陕西省科技研究发展计划(2012KCT-16)
【分类号】：R743.3

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