[1]高继英,石代乐,王 刚.长链非编码RNA NORAD过表达通过靶向调控miR-132-5p/Bcl-2改善MPP+诱导的帕金森细胞模型损伤实验研究[J].陕西医学杂志,2023,52(4):363-368,384.[doi:DOI:10.3969/j.issn.1000-7377.2023.04.001]
　GAO Jiying,SHI Daile,WANG Gang.LncRNA NORAD overexpression improves MPP+ induced Parkinson cell model injury by targeting miR-132-5p/Bcl-2[J].,2023,52(4):363-368,384.[doi:DOI:10.3969/j.issn.1000-7377.2023.04.001]
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长链非编码RNA NORAD过表达通过靶向调控miR-132-5p/Bcl-2改善MPP+诱导的帕金森细胞模型损伤实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年4期
页码:
363-368,384
栏目:
基础研究
出版日期:
2023-04-05

文章信息/Info

Title:
LncRNA NORAD overexpression improves MPP+ induced Parkinson cell model injury by targeting miR-132-5p/Bcl-2
作者:
高继英1; 石代乐1; 王 刚2
(1.河北北方学院附属第一医院神经外科,河北 张家口075000; 2.河北北方学院附属第一医院神经内科,河北 张家口075000)
Author(s):
GAO Jiying; SHI Daile; WANG Gang
(Department of Neurosurgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)
关键词:
帕金森病;  MN9D细胞;  长链非编码RNA NORAD;  细胞凋亡;  氧化应激;  炎症
Keywords:
Parkinson's disease;  MN9D cell;  Long non-coding RNA NORAD;  Apoptosis;  Oxidative stress;  Inflammation
分类号:
R 745.1
DOI:
DOI:10.3969/j.issn.1000-7377.2023.04.001
文献标志码:
A
摘要:
目的:探讨长链非编码RNA(lncRNA)NORAD对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森(PD)细胞模型损伤的影响,及其潜在分子作用机制。方法:3 mmol/L的MPP+处理MN9D细胞建立PD细胞损伤模型,MN9D细胞转染lncRNA NORAD过表达载体、miR-132-5p mimics、miR-132-5p inhibitor、Bcl-2 siRNA及相应对照,qRT-PCR检测细胞中lncRNA NORAD、miR-132-5p和Bcl-2 mRNA的表达,Western blot检测细胞中Bcl-2 蛋白表达,流式细胞仪检测细胞凋亡,试剂盒法检测细胞培养液中丙二醛(MDA)、超氧化物歧化酶(SOD)和肿瘤坏死因子-α(TNF-α)、白介素1β(IL-1β)的水平,双荧光素酶实验验证lncRNA NORAD、miR-132-5p和Bcl-2之间的靶向关系。结果:MPP+诱导MN9D细胞中lncRNA NORAD、Bcl-2 mRNA和蛋白表达降低,诱导细胞中miR-132-5p表达升高; 过表达lncRNA NORAD能够减轻MPP+诱导的MN9D细胞凋亡、氧化应激和炎症,保护MN9D细胞; lncRNA NORAD与miR-132-5p靶向结合,过表达lncRNA NORAD抑制MN9D细胞中miR-132-5p的表达; miR-132-5p mimics共转染能够逆转lncRNA NORAD过表达对MPP+诱导的MN9D细胞损伤的保护作用; miR-132-5p与Bcl-2的3'UTR靶向结合,转染miR-132-5p inhibitor下调MN9D细胞中miR-132-5p的表达,而促进Bcl-2 mRNA和蛋白表达; 转染miR-132-5p inhibitor能够减轻MPP+诱导的MN9D细胞凋亡、氧化应激和炎症,保护MN9D细胞,而共转染Bcl-2 siRNA则能逆转miR-132-5p inhibitor对MPP+诱导的MN9D细胞损伤的保护作用。结论:lncRNA NORAD过表达可通过靶向调控miR-132-5p/Bcl-2,减轻MPP+诱导的MN9D细胞凋亡、氧化应激和炎症,对MN9D细胞损伤具有保护作用。
Abstract:
Objective:To investigate the effect of lncRNA NORAD on MPP+ induced Parkinson's disease(PD)cell model injury and its potential molecular mechanism.Methods:The damage model of PD cells was established by treating MN9D cells with 3 mmol/L MPP+.MN9D cells were transfected with lncRNA NORAD overexpression vector,miR-132-5p mimics,miR-132-5p inhibitor,Bcl-2 siRNA and corresponding control.The expressions of lncRNA NORAD,miR-132-5p and Bcl-2 mRNA in the cells were detected by qRT-PCR.The protein expression of Bcl-2 in the cells was detected by Western blot.The apoptosis was detected by flow cytometry.The levels of MDA,SOD,TNF-α and IL-1 in the cell culture medium were detected by the kit method.The targeting relationship between lncRNA NORAD,miR-132-5p and Bcl-2 was verified by the dual-luciferase assay.Results:MPP+ decreased lncRNA NORAD,Bcl-2 mRNA and protein expression,and increased miR-132-5p expression in MN9D cells.Overexpression lncRNA NORAD could reduce MPP+ induced apoptosis,oxidative stress and inflammation of MN9D cells,and protect MN9D cells.LncRNA NORAD targeted miR-132-5p,and overexpression of lncRNA NORAD inhibited the expression of miR-132-5p in MN9D cells.MiR-132-5p mimics co-transfection could reverse the protective effect of lncRNA NORAD overexpression on MPP+ induced MN9D cell injury.MiR-132-5p was targeted to bind with the 3'UTR of Bcl-2,and transfected with miR-132-5p inhibitor to down-regulate the expression of miR-132-5p in MN9D cells,while promoting the expression of Bcl-2 mRNA and protein.Transfection with miR-132-5p inhibitor could reduce MPP+ induced apoptosis,oxidative stress and inflammation of MN9D cells,and protect MN9D cells,while co-transfection with Bcl-2 siRNA could reverse the protective effect of miR-132-5p inhibitor on MPP+ induced damage of MN9D cells.Conclusion:LncRNA NORAD overexpression could reduce MPP+ induced apoptosis,oxidative stress and inflammation of MN9D cells through targeted regulation of miR-132-5p/Bcl-2 axis,and had a protective effect on MN9D cell damage.

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备注/Memo

备注/Memo:
基金项目:河北省医学科学研究课题(20220029)
更新日期/Last Update: 2023-04-06