miR-138调控Akt/PI3K信号通路对缺氧缺糖诱导大鼠皮层神经细胞损伤的影响  

Effects of miR-138 Regulation Akt/PI3K Signaling Pathway on Cortical Neuronal Damage Induced by Hypoxia and Hypoglycemia in Rats

作　　者：李敏 王红艳 邵明阳 LI Min;WANG Hongyan;SHAO Mingyang(Zaozhuang Hospital,Zaozhuang Mining Group,Zaozhuang 277100,Shandong,China)

机构地区：[1]枣庄矿业集团枣庄医院,山东枣庄277100

出　　处：《中西医结合心脑血管病杂志》2021年第21期3685-3690,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的探讨miR-138调控蛋白激酶B(Akt)/磷脂酰肌醇3激酶(PI3K)信号通路对缺氧缺糖诱导大鼠皮层神经细胞损伤的影响。方法构建大鼠皮层神经细胞缺氧缺糖模型,在细胞中转染miR-138 mimics,荧光定量聚合酶链式反应(qRT-PCR)方法测定miR-138表达变化,四甲基偶氮唑盐(MTT)比色法检测细胞活性,试剂盒检测乳酸脱氢酶(LDH)释放率,流式细胞术检测细胞凋亡率,蛋白免疫印迹法(Western Blot)检测细胞中Bax、Bcl-2、磷酸化蛋白激酶B(p-Akt)、磷酸化磷脂酰肌醇3激酶(p-PI3K)蛋白表达。用Akt/PI3K信号抑制剂处理上调miR-138的大鼠皮层神经细胞,经缺氧缺糖处理后,检测细胞活性、LDH释放率和细胞凋亡率。结果缺氧缺糖大鼠皮层神经细胞中miR-138表达水平下降,细胞活性降低,LDH释放率和细胞凋亡率升高,Bax蛋白表达增多,Bcl-2、p-Akt、p-PI3K蛋白表达减少,差异均有统计学意义(P<0.05)。转染miR-138 mimics可以提高缺氧缺糖的大鼠皮层神经细胞中miR-138表达水平,提高细胞活性,降低LDH释放率并减少细胞凋亡,减少Bax蛋白表达,促进Bcl-2、p-Akt、p-PI3K蛋白表达,差异均有统计学意义(P<0.05)。Akt/PI3K信号抑制剂可以逆转上调miR-138对缺氧缺糖大鼠皮层神经细胞活性、LDH释放率和凋亡作用,差异均有统计学意义(P<0.05)。结论上调miR-138通过激活Akt/PI3K信号通路减轻缺氧缺糖诱导大鼠皮层神经细胞损伤,减少细胞凋亡。Objective To investigate the effect of miR-138 regulation serine-threonine kinase(Akt)/phosphoinositide 3-kinase(PI3K)signaling pathway on cortical nerve cell injury induced by hypoxia and hypoglycemia in rats.Methods The rat model with hypoxia and hypoglycemia in cortical neurons was constructed,miR-138 mimics were transfected in the cells,miR-138 expression was measured by quantitative real-time polymerase chain reaction(qRT-PCR)method.Cell activity was detected by methyl thiazolyl tetrazolium(MTT).Lactate dehydrogenase(LDH)release rate was detected by kit,and apoptosis was detected by flow cytometry.Bax,Bcl-2,p-Akt,and p-PI3K protein expressions were detected by Western Blot.Rat cortical neurons up-regulated with miR-138 were treated with Akt/PI3K signal inhibitors.After hypoxia and hypoglycemia treatment,Cell activity.LDH release rate,and apoptosis were measured.Results The expression of miR-138 in hypoxic and hypoglycemic rat cortical neurons decreased.Cell activity decreased.LDH release rate and apoptosis rate increased,Bax protein expression increased.Bcl-2,p-Akt,and p-PI3K proteins expression reduced(P<0.05).Transfection of miR-138 mimics could increase the expression level of miR-138 in rat hypoxic and hypoglycemic cortical neurons,and increase cell activity,reduce LDH release rate and apoptosis,reduce Bax protein expression,and promote Bcl-2,p-Akt,p-PI3K protein expression(P<0.05).Akt/PI3K signal inhibitors could reverse the up-regulation of miR-138 on cortical neuronal activity,LDH release rate,and apoptosis in hypoxic and hypoglycemic rats(P<0.05).Conclusion Up-regulating miR-138 by activating Akt/PI3K signaling pathway could attenuate hypoxia and hypoglycemic-induced damage in rat cortical neurons and reduces apoptosis.

关 键 词：神经细胞损伤 缺氧缺糖诱导 miR-138 Akt/PI3K信号通路 实验研究 

分 类 号：R73[医药卫生—肿瘤]