miR-185抑制VEGFA导致抑郁症大鼠海马神经元损伤  

MicroRNA-185 Induces Hippocampal Neuron Damage in Depression Rats by Inhibiting VEGFA

作　　者：孙玉涛[1] 王磊[2] 贾翠娜 徐阳 黄艳艳 沈振明[3] 赵安全[5] SUN Yutao;WANG Lei;JIA Cuina;XU Yang;HUANG Yanyan;SHEN Zhenming;ZHAO Anquan(Psychological Ward,Tangshan No.5 Hospital,Tangshan,He­bei,063000,China;Department cf Fourth Ward of Inpatient,Tangshan No.5 Hospital,Tangshan,He­bei,063000,China;Department of Rehabilitation,Tangshan No.5 Hospital,Tangshan,He­bei,063000,China;De­partment of Second Ward of Inpatient,Tangshan No.5 Hospital,Tangshan,He­bei,063000,China;Director Office,Tangshan No.5 Hospital,Tangshan,He­bei,063000,China)

机构地区：[1]唐山市第五医院心理病区,河北省唐山市063000 [2]唐山市第五医院住院部四病区,河北省唐山市063000 [3]唐山市第五医院康复科,河北省唐山市063000 [4]唐山市第五医院住院部二病区,河北省唐山市063000 [5]唐山市第五医院院长办,河北省唐山市063000

出　　处：《医学分子生物学杂志》2021年第4期273-279,共7页Journal of Medical Molecular Biology

基　　金：河北省2021年度医学科学研究课题计划(No.20211102)。

摘　　要：目的探索miR-185对抑郁症模型大鼠海马区神经细胞的影响.方法采用实施慢性不可预知的轻度压力(chronic unpredictable mild stress,CUMS)模拟大鼠抑郁症模型.SD大鼠40只随机分为2组(n=20):对照组(Control)、模型组(Model).记录两组大鼠体重、检测大鼠糖水偏好及通过旷场实验对大鼠行为学进行评分.Western印迹检测海马组织血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、Bax、Bcl-2和Ki67的表达水平;qRT-PCR检测两组大鼠海马组织和原代海马神经元细胞的miR-185和VEGFA的mRNA水平.原代分离大鼠海马区神经细胞.荧光素酶报告实验验证miR-185与VEGFA靶向关系;细胞分为4组:健康组(Con-trol,从健康组大鼠分离)、模型组(Model,从模型组大鼠分离)、miR-185 inhibitor(从模型组大鼠分离,并转染miR-185 inhibitor)组和miR-185 mock组(从模型组大鼠分离,并转染miR-185 mock);分别检测各组细胞的细胞活力(CCK-8法)、细胞凋亡(流式细胞术)以及相关蛋白的表达水平.结果与对照组相比,模型组海马组织miR-185表达水平升高(P<0.01),而VEGFA表达水平下降(P<0.01);且除Bax增加外,VEGFA、BDNF、Bcl-2和Ki67的表达水平较对照组减少(P<0.01);与模型组相比,miR-185 inhib-itor抑制miR-185表达,增加VEGFA表达,miR-185 mimic有相反的效果.生物信息学预测和荧光素酶报告实验证明miR-185靶向结合到VEGFA 3′-未翻译区域(3′-UTR).与对照组相比,模型组和miR-185 mock组细胞活力下降,细胞凋亡率增加,且除Bax增加外,VEGFA、BDNF、Bcl-2和Ki67的表达水平较对照组减少(P<0.01);与模型组相比,miR-185 inhibitor增加细胞活力,降低凋亡率,减少Bax/Bcl2比率,增加VEGFA、BDNF和Ki67的表达水平.结论miR-185通过下调VEGFA导致抑郁症大鼠海马区神经细胞损伤.Objective To explore the effect of microRNA-185(miR-185)on neuron cells in the hippocampus of depression rats.Methods The chronic unpredictable mild stress(CUMS)model of SD rat was established.Rats were randomly divided into two groups(n=20):control group and model group.The weight of rats was recorded,and the sucrose preference test and the open-field test were performed.The expression levels of VEGFA,BDNF,Bax,Bcl-2 and Ki67 in hippocampus were detected by Western blotting,and the mRNA levels of miR-185 and VEGFA in hippocampus or primary cultured hippocampal neuron cells by qRT-PCR.The target relationship between miR-185 and VEGFA was verified by the luciferase reporter assays.Cells were divided into 4 groups:control,model,miR-185 inhibitor and miR-185 mock group.The viability and apoptosis of cells were measured by CCK-8 and flow cytometry,respectively,and the levels of related pro­teins were determined.Results The expression level of miR-185 was significantly increased and that of VEGFA significantly decreased in the hippocampus of the model group compared with the control group(P<0.01 for both).Except for the increased expression level of Bax,the expression levels of VEGFA,BDNF,Bcl-2 and Ki67 were significantly reduced in the model group compared with the control group(P<0.01).The miR-185 inhibitor inhibited the miR-185 expression and promoted the VEGFA expression,while the opposite effect was observed in the miR-185 mock group.Bioinformatics prediction and luciferase reporter assays demonstrated that miR-185 targeted the 3'-untranslated region(3'-UTR)of VEGFA.The cell viability was significantly decreased,and the cell apoptosis rate significantly increased in the model group and miR-185 mock group compared with the control group(P<0.01 for all).Except for the increased expression level of Bax,the ex­pression levels of VEGFA,BDNF,Bcl-2 and Ki67 were significantly reduced in the model group and miR-185 mock group compared with the control group(P<0.01).The miR-185 inhibitor in­creased the cell viability,re

关 键 词：抑郁症 miR-185 血管内皮生长因子A 

分 类 号：R749.1[医药卫生—神经病学与精神病学]