miR-150-5p通过调控DDR1表达对缺氧缺糖诱导的大鼠皮质神经细胞损伤的影响

Effects of miR-150-5p on oxygen-glucose deprivation-induced damage to rat cortical nerve cells by regulating DDR1 expression

目的:探讨miR-150-5p对缺氧缺糖(OGD)诱导的大鼠皮质神经细胞损伤的影响及分子机制。方法:设置OGD组、正常对照(Con)组、miR-NC组、miR-150-5p组、anti-miR-NC组、anti-miR-150-5p组、OGD+miR-NC组、OGD+miR-150-5p组、OGD+si-NC组、OGD+si-DDR1组、OGD+miR-150-5p+pcDNA组、OGD+miR-150-5p+pcDNA-DDR1组。RT-qPCR检测miR-150-5p和DDR1 mRNA表达;Western blot检测盘状结构域受体1(DDR1)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达;MTT检测细胞存活率;乳酸脱氢酶(LDH)试剂盒检测LDH释放率;流式细胞术检测细胞凋亡;双荧光素酶报告实验验证miR-150-5p和DDR1靶向关系。结果:OGD诱导的大鼠皮质神经细胞中miR-150-5p呈低表达,DDR1呈高表达,细胞存活率降低,LDH释放率升高,细胞凋亡率升高,Bcl-2表达降低,Bax表达升高(P<0.05)。过表达miR-150-5p或抑制DDR1表达可促进细胞存活,抑制LDH释放和细胞凋亡。miR-150-5p靶向调控DDR1,DDR1过表达逆转miR-150-5p过表达对OGD诱导的大鼠皮质神经细胞损伤的保护作用。结论:过表达miR-150-5p通过下调DDR1保护缺氧缺糖诱导的大鼠皮质神经细胞损伤。

Objective:To investigate effect of miR-150-5 p on oxygen-glucose deprivation(OGD)induced injury of rat cortical neuronal cells and its molecular mechanism.Methods:Set up OGD group,normal control(Con)group,miR-NC group,miR-150-5 p group,anti-miR-NC group,anti-miR-150-5 p group,OGD+miR-NC group,OGD+miR-150-5 p group,OGD+si-NC group,OGD+siDDR1 group,OGD+miR-150-5 p +pcDNA group,OGD+miR-150-5 p+pcDNA-DDR1 group. RT-qPCR to detect expressions of miR-150-5 p and DDR1 mRNA. Western blot to detect expressions of discoid domain receptor 1(DDR1),B-cell lymphoma/leukemia-2(Bcl-2),and Bcl-2 related X protein(Bax)proteins. MTT to detect cell survival rate. Lactate dehydrogenase(LDH)kit to detect LDH release rate. Flow cytometry was used to detect apoptosis. Dual luciferase report experiment to verify targeting relationship between miR-150-5 p and DDR1.Results:Low expression of miR-150-5 p and high expression of DDR1 were in OGD induced rat cortical neurons,cell survival was decreased,LDH release was increased,apoptosis was increased,Bcl-2 expression was decreased,and Bax expression was increased(P<0.05). Overexpression of miR-150-5 p or inhibition of DDR1 expression promoted cell survival,inhibited LDH release and cell apoptosis. miR-150-5 p targeting DDR1,and overexpression of DDR1 reversed protective effect of miR-150-5 p overexpression on OGD-induced rat cortical neuronal injury.Conclusion:Overexpression of miR-150-5 p protects rat cortical neurons from injury induced by oxygen-glucose deprivation by down-regulating DDR1.