摘要目的:研究微RNA-873-5p(miR-873-5p)靶向调控电压依赖性阴离子通道1(VDAC1)对大鼠癫痫样神经细胞的影响及作用机制。方法:通过无镁培养基诱导构建大鼠癫痫样海马神经细胞模型，将大鼠神经细胞分为对照组、模型组、miR-con组及miR-873-5p组。对照组采用正常培养大鼠神经细胞；模型组采用低镁细胞外液培养大鼠神经细胞；miR-con组采用低镁细胞外液培养大鼠神经细胞并转染miR-con；miR-873-5p组采用低镁细胞外液培养大鼠神经细胞并转染miR-873-5p。采用荧光定量聚合酶链式反应(qRT-PCR)检测各组神经细胞miR-873-5p及VDAC1 mRNA表达；采用Western blot技术检测神经细胞中VDAC1、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸天冬氨酸蛋白水解酶3(Cleaved caspase-3)含量；采用DCFH-DA荧光探针检测神经细胞活性氧(ROS)水平；采用硫代巴比妥酸法检测神经细胞丙二醛(MDA)含量；采用酶联免疫吸附实验(ELISA)检测神经细胞谷胱甘肽(GSH)含量；采用流式细胞术检测各组神经细胞凋亡情况；采用双荧光素酶报告基因法验证miR-873-5p对VDAC1的靶向调控作用。结果:与对照组比较，模型组神经细胞中miR-873-5p表达显著降低( P<0.05)，VDAC1表达显著升高( P<0.05)，Bcl-2表达下调( P<0.05)，Bax及Cleaved caspase-3表达上调( P<0.05)，GSH及MDA含量显著降低( P<0.05)，而ROS水平显著升高( P<0.05)，细胞凋亡率显著升高( P<0.05)；与miR-con组比较，miR-873-5p组神经细胞中Bcl-2表达明显上调( P<0.05)，Bax及Cleaved caspase-3表达明显下调( P<0.05)，GSH及MDA含量显著升高( P<0.05)，ROS水平显著降低( P<0.05)，细胞凋亡率明显降低( P<0.05)；双荧光素酶报告基因实验表明miR-873-5p能抑制VDAC1表达( P<0.05)。 结论:miR-873-5p能通过负向调控靶基因VDAC1表达而减少神经细胞凋亡，同时还能抑制神经细胞氧化应激反应，对受损神经细胞具有保护作用。

abstractsObjective:To explore the regulatory effect of miR-873-5p micro-RNA targeting voltage-dependent anion channel protein 1 (VDAC1) in neurons and its mechanism.Methods:Murine nerve cells were randomly divided in vitro into a control group, a model group, a mimetic negative carrier (miR-con) group and an miR-873-5p group. The epileptiform hippocampal nerve cell model was induced in all of the cells except those in the control group using magnesium-free medium. The control group was normally cultured, while the miR-con and miR-873-5p groups were transfected with miR control and miR-873-5p RNA respectively. Real-time fluorescent quantitative polymerase chain reactions were used to detect the expression of miR-873-5p and VDAC1 mRNA. Western blotting was employed to detect VDAC1, B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax) and cleared caspase-3 in the neurons. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) were measured using the DCFH-DA fluorescent probe, the thiobarbituric acid method and enzyme-linked immunosorbent assay respectively. Any apoptosis was detected using flow cytometry, while the targeting of miR-873-5p on VDAC1 was verified using the double fluorescence zymase reporter gene method.Results:Compared with the control group, a significant decrease in the average expression of miR-873-5p, Bcl-2 and in GSH and MDA levels was observed in the model group, but there was a significant increase in the average level of VDAC1, Bax, cleaved caspase-3 and ROS and in the rate of apoptosis. Compared with the miR-con group, a significant decrease in the average expression of Bax, cleaved caspase-3, ROS and in the apoptosis rate was observed in the miR-873-5p group, but there was a significant increase in the average level of Bcl-2, GSH and MDA. Moreover, it was verified that miR-873-5p reduced the expression of VDAC1.Conclusion:miR-873-5p protects damaged neurons by inhibiting their apoptosis through negatively regulating the target gene VDAC1 and the oxidative stress response.