miR-10a通过BDNF信号通路对精神分裂症大鼠海马神经元细胞的作用研究

Effects of miR-10aon hippocampal neurons ofschizophrenia rats through BDNF signal path⁃way

目的:探究miR-10a对精神分裂症(SZ)大鼠海马神经元细胞的作用及其可能的机制。方法:30只大鼠随机分为空白对照组、生理盐水组和SZ组,每组10只。应用MK-801建立SZ大鼠模型。Morris水迷宫实验检测学习记忆能力;旷场实验检测大鼠自主行为与紧张度;实时荧光定量聚合酶链反应(qPCR)和Western blot检测miR-10a、脑源性神经营养因子(BDNF)通路相关基因的表达;荧光素酶报告基因实验验证miR-10a与BDNF之间的靶向结合作用;CCK-8检测海马神经元细胞活力;流式细胞术检测海马神经元细胞周期及凋亡情况。结果:与生理盐水组相比,SZ组大鼠逃跑潜伏期、移动距离、移动速度和垂直次数均显著增加(P<0.05);与空白对照组相比,SZ组大鼠海马BDNF通路被抑制,miR-10a表达显著升高(P<0.05);BDNF为miR-10a的靶基因;与NC组相比,miR-10a mimic和K252a显著抑制SZ大鼠海马神经元细胞BDNF通路和细胞活力(P<0.05),阻滞细胞周期进程,促进细胞凋亡,miR-10a inhibitor则作用相反;与miR-10a inhibitor组相比,miR-10a inhibitor+K252a处理显著抑制BDNF通路活化和细胞活力(P<0.05),阻滞细胞周期进程,促进细胞凋亡。结论:miR-10a通过BDNF信号通路抑制SZ大鼠海马神经元细胞增殖,促进细胞凋亡。

Objective:To explore the effect and its possible mechanismof miR-10a on hippocampal neurons of schizophrenic(SZ)rats.Methods:A total of 30 rats were randomly divided into blank control group,saline group and SZ group,with 10 rats in each group.SZ rat model was established by MK-801.Morris water maze test was used to detect learning and memory ability.Open field test was used to detect autonomous behavior and tension in rats.qPCR and Western blot were used to detect the expression of genes related to miR-10a and brain-derived neurotrophic factor(BDNF)pathway.Luciferase reporter gene assay verified the targeted binding between miR-10a and BDNF.CCK-8 detected the viability of hippocampal neurons.The cell cycle and apoptosis of hippocampal neurons were detected by flow cytometry.Results:Compared with the normal saline group,the escape latency,moving distance,moving speed and vertical times were significantly increased in the SZ group(P<0.05).Compared with the blank control group,the BDNF pathway in the hippocampus was inhibited and the expression of miR-10a was significantly increased in the SZ group(P<0.05).BDNF was the target gene of miR-10a.Compared with NC group,miR-10a mimic and K252a significantly inhibited BDNF pathway and cell viabilityin hippocampal neurons of SZ rats(P<0.05),blocked cell cycle progression and promoted apoptosis,while miR-10a inhibitor had the opposite effect.Compared with miR-10a inhibitor group,miR-10a inhibitor+K252a significantly inhibited BDNF pathway activation and cell viability(P<0.05),blocked cell cycle progression and promoted cell apoptosis.Conclusion:MiR-10a can inhibit the proliferation of hippocampal neurons of SZ rats and promote apoptosis through BDNF signal pathway.