过表达lncRNA TUG1缓解帕金森病小鼠模型神经元线粒体自噬缺陷和神经元损伤的分子机制研究  

The Molecular Mechanisms of lncRNA TUG1 Overexpression in Alleviating Parkinson's Disease Mice Model Neuronal Mitophagy Defects and Neuronal Injury

作　　者：赵阳 张文君 方敬献[1] ZHAO Yang;ZHANG Wen-jun;FANG Jing-xian(Department of Neurology,the First People's Hospital of Nanyang City,Nanyang 473000,China)

机构地区：[1]南阳市第一人民医院神经内一科,南阳473000

出　　处：《中国临床神经科学》2024年第5期489-495,共7页Chinese Journal of Clinical Neurosciences

基　　金：河南省科技计划发展项目(编号:202102310407)。

摘　　要：目的探讨过表达lncRNA TUG1保护帕金森病(PD)模型小鼠大脑黑质神经元损伤的分子机制。方法40只雄性C57/BL6小鼠随机分为对照组、模型组(建立PD模型)、模型+Lentiv-lncRNA TUG1-OE组(PD模型建立后经尾静脉注射慢病毒颗粒介导过表达lncRNA TUG1)、模型+Lentiv组(PD模型建立后经尾静脉注射空载慢病毒颗粒,每组n=10。取小鼠大脑黑质组织。qPCR法测定小鼠大脑黑质组织中lncRNA TUG1和PGC-1αmRNA的表达水平。Westernblot法测定其中PGC-1α,自噬相关蛋白LC3-Ⅱ/Ⅰ比值,Beclin-1和P62表达水平,以及线粒体自噬相关蛋白PINK1和Parkin的表达水平。荧光探针法测定小鼠大脑黑质中ROS水平。ELISA法测定还原型GSH/GSSG比率、ATP水平、IL-6和IL-1β水平。结果与对照组比较,模型组小鼠大脑黑质中lncRNA TUG1和PGC-1α蛋白及mRNA表达水平、GSH/GSSH比值、ATP、PINK1、Parkin和P62蛋白相对表达水平降低(均P<0.05);ROS相对水平,IL-6和IL-1β水平,LC3-Ⅱ/Ⅰ比值以及Beclin-1相对表达水平均升高(均P<0.05)。与模型组比较,模型+Lentiv-LncRNA TUG1-OE组部分逆转了上述检测指标水平(均P<0.05);而模型+Lentiv组上述检测指标差异无显著性(均P<0.05)。结论过表达lncRNA TUG1可缓解PD模型小鼠神经元线粒体自噬功能障碍,可能对神经元损伤发挥保护功效。Aim To explore the molecular mechanisms of lncRNA TUG1 overexpression in protecting neuronal injury in Parkinson's disease(PD)mice model substantia nigra.Method Forty male C57/BL6 mice were randomly divided into a control group,a model group,a model+Lentiv-lncRNA TUG1-OE group and a model+Lentiv group.PD mouse model was constructed in the latter three groups.In the model+Lentiv-lncRNA TUG1-OE group,lentivirus particles were injected into mice tail vein to induce overexpression of lncRNA TUG1.In the model+Lentiv group,vector lentivirus particles were injected into mice tail vein.The brain substantia nigra tissues of mice were collected.qPCR analysis was used to measure the expression levels of lncRNA TUG1 and PGC-1αmRNA in substantia nigra of mice brain.Western blot was used to determine the expression levels of PGC-1α,LC3-Ⅱ/Ⅰratio,Beclin-1 and p62,as well as PINK1 and Parkin.The ROS levels were determined by fluorescence probe.ELISA was used to measure the GSH/GSSG ratio,ATP levels,IL-6 and IL-1βlevels.Results Compared with the control group,the expression levels of lncRNA TUG1,PGC-1αmRNA and protein,GSH/GSSH ratio,ATP levels,the relative expression levels of PINK1,Parkin and p62 in substantia nigra of the model group mice decreased(P<0.05).The relative levels of ROS,the levels of IL-6 and IL-1β,the relative expression levels of LC3-Ⅱ/Ⅰratio and Beclin-1 increased(P<0.05).Compared with the model group,the model+Lentiv-lncRNA TUG1-OE group partially reversed the levels of the above detected indicators(P<0.05).However,there was no significant difference of the above indicators in model+Lentiv group(P<0.05).Conclusion Overexpression of lncRNA TUG1 can alleviate mitophagy dysfunction in PD model mice,and may play a protective role in neuronal injury.

关 键 词：帕金森病 长链非编码RNA牛磺酸上调基因1 过氧化物酶体增殖物激活受体-γ共激活因子1α 线粒体自噬 自噬 

分 类 号：Q189[生物学—神经生物学]