【摘要】：目的:探討p38α蛋白在腦缺血后2 h降低的原因以及p38α蛋白水平表達是否受調(diào)于microRNA-128。方法:首先通過qPCR和Western blotting來檢測p38αmRNA和蛋白的表達水平,其次通過生物信息學軟件預測出p38α的表達可能受調(diào)于microRNA-128,最后進行細胞水平驗證,使用攜帶有microRNA-128序列質(zhì)粒的慢病毒載體、攜帶有microRNA-128反義序列質(zhì)粒的慢病毒載體以及相應空質(zhì)粒的慢病毒載體轉(zhuǎn)染SH-SY5Y細胞,觀察p38α蛋白的表達情況。結(jié)果:小鼠腦缺血后2 h p38αmRNA水平無降低,但其蛋白水平卻明顯下降,microRNA-128的表達水平明顯升高;轉(zhuǎn)染實驗發(fā)現(xiàn),轉(zhuǎn)染microRNA-128序列質(zhì)粒的慢病毒載體的細胞p38α蛋白水平明顯降低,轉(zhuǎn)染microRNA-128反義序列質(zhì)粒的慢病毒載體的細胞p38α蛋白水平明顯升高。結(jié)論:腦缺血后p38α蛋白的表達水平下降并不是因為p38αmRNA水平的降低,microRNA-128可以調(diào)控p38α蛋白的表達,可能是通過結(jié)合p38α的mRNA從而抑制其蛋白的翻譯表達。
[Abstract]:Objective: to investigate the causes of the decrease of p38 偽 protein at 2 h after cerebral ischemia and whether the expression of p38 偽 protein is regulated in microRNA-128.. Methods: firstly, the expression of p38 偽 mRNA and protein was detected by qPCR and Western blotting, and then the expression of p38 偽 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 偽 protein. Results: at 2 h after cerebral ischemia, the level of p38 偽 mRNA did not decrease, but the protein level decreased significantly, and the expression of microRNA-128 increased significantly. The p38 偽 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 偽 protein expression after cerebral ischemia is not due to the decrease of p38 偽 mRNA level. The expression of p38 偽 protein can be regulated by microRNA-128, which may inhibit the translation of p38 偽 protein by binding p38 偽 mRNA.
【作者單位】： 西安交通大學醫(yī)學部第三附屬醫(yī)院神經(jīng)外科;
【基金】：陜西省科技研究發(fā)展計劃(2012KCT-16)
【分類號】：R743.3

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