长链非编码RNA 细胞骨架调节RNA靶向微小RNA-1246对帕金森病模型细胞损伤的保护作用

doi:10.16098/j.issn.0529-1356.2022.05.004

解剖学报 ›› 2022, Vol. 53 ›› Issue (5): 563-570.doi: 10.16098/j.issn.0529-1356.2022.05.004

长链非编码RNA 细胞骨架调节RNA靶向微小RNA-1246对帕金森病模型细胞损伤的保护作用

李薇^1* 王丽² 汪志华³ 刘庆春¹ 韩荣胜⁴

1.青海省第五人民医院神经内科，西宁 810000; 2.青海省第五人民医院内分泌科，西宁 810000; 3.青海心脑血管病专科医院影像科，西宁 810012; 4.青海省第五人民医院心内科，西宁 810000

收稿日期:2020-12-11 修回日期:2021-05-13 出版日期:2022-10-06 发布日期:2022-10-06
通讯作者: 李薇 E-mail:vks081@163.com

Protective effect of long non-coding RNA cytoskeleton regulatory RNA targeting microRNA-1246 on cell damage in Parkinson’s disease models

LI Wei^1* WANG Li² WANG Zhi-hua³ LIU Qing-chun¹ HAN Rong-sheng⁴

1.Department of Neurology, the Fifth People’s Hospital of Qinghai Province, Xining 810000, China; 2.Department of Endocrinology, the Fifth People’s Hospital of Qinghai Province, Xining 810000, China; 3.Department of Imaging, Qinghai Cardiovascular and Cerebrovascular Disease Hospital, Xining 810012, China;4.Department of Cardiology, the Fifth People’s Hospital of Qinghai Province, Xining 810000, China

Received:2020-12-11 Revised:2021-05-13 Online:2022-10-06 Published:2022-10-06
Contact: LI Wei E-mail:vks081@163.com

摘要/Abstract

摘要：

目的探讨长链非编码RNA（lncRNA）细胞骨架调节RNA（CYTOR）靶向微小RNA（miR）-1246对帕金森病（PD）模型细胞损伤的影响。方法采用100 μmol/L 1-甲基-4-苯基-吡啶离子（MPP+）诱导SK-N-SH细胞建立PD细胞模型。Real-time PCR检测CYTOR和miR-1246表达。在SK-N-SH细胞中转染pcDNA-CYTOR或anti-miR-1246，或共转染pcDNA-CYTOR和miR-1246模拟物，经MPP+处理后，采用流式细胞术分析细胞凋亡，试剂盒检测丙二醛（MDA）含量及谷胱甘肽过氧化物酶（GSH）活性。双荧光素酶报告基因实验分析CYTOR和miR-1246靶向关系。结果 PD细胞模型中CYTOR表达量降低（P<0.05），miR-1246表达量升高（P<0.05）。过表达CYTOR或抑制miR-1246表达均可降低模型细胞凋亡率和MDA含量（P<0.05），并增加GSH活性（P<0.05）。MiR-1246是CYTOR的靶基因，CYTOR负性调控miR-1246表达。上调miR-1246可逆转CYTOR过表达对模型细胞凋亡和氧化损伤的影响（P<0.05）。结论 LncRNA CYTOR靶向miR-1246可减轻PD模型细胞凋亡和氧化损伤。

关键词: 长链非编码RNA, , 细胞骨架调节RNA, 微小RNA-1246, 氧化损伤, 流式细胞术, 帕金森病细胞模型

Abstract:

Objective To study the effect of long non-coding RNA (lncRNA) cytoskeleton regulatory RNA (CYTOR) targeting microRNA (miR)-1246 on cell damage in Parkinson’s disease (PD) models. Methods SK-N-SH cells were exposed to 100 μmol/L 1-methyl-4-phenylpyridinium (MPP+) to establish a PD cell model in vitro. The levels of CYTOR and miR-1246 were determined by Real-time PCR. pcDNA-CYTOR or anti-miR-1246, or co-transfect pcDNA-CYTOR and miR-1246 mimics were transfected into SK-N-SH cells, repectively. After treated with MPP+, flow cytometry was used to assess cell apoptosis, and commercial kits was used to monitor malondialdehyde (MDA) content and glutathione peroxidase (GSH) activity. The interaction between CYTOR and miR-1246 was verified by dual-luciferase reporter assay. Results CYTOR reduced (P<0.05), while miR-1246 increased (P<0.05) in PD model cells. CYTOR overexpression or miR-1246 inhibition could reduce the apoptosis rate and MDA content of model cells (P<0.05), and increase GSH activity (P<0.05). MiR-1246 was the target gene of CYTOR, and CYTOR negatively regulated miR-1246 expression. Up-regulation of miR-1246 could reverse the effect of CYTOR overexpression on apoptosis and oxidative damage of model cells (P<0.05). Conclusion LncRNA CYTOR can reduce cell apoptosis and oxidative damage in PD models by targeting miR-1246.

Key words: Long non-coding RNA, Cytoskeleton regulatory RNA, MicroRNA-1246, Oxidative damage, Flow cytometry, Parkinson’s disease cell model

中图分类号:

R742

李薇王丽汪志华刘庆春韩荣胜. 长链非编码RNA 细胞骨架调节RNA靶向微小RNA-1246对帕金森病模型细胞损伤的保护作用[J]. 解剖学报, 2022, 53(5): 563-570.

LI Wei WANG Li WANG Zhi-hua LIU Qing-chun HAN Rong-sheng . Protective effect of long non-coding RNA cytoskeleton regulatory RNA targeting microRNA-1246 on cell damage in Parkinson’s disease models[J]. Acta Anatomica Sinica, 2022, 53(5): 563-570.

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长链非编码RNA 细胞骨架调节RNA靶向微小RNA-1246对帕金森病模型细胞损伤的保护作用

Protective effect of long non-coding RNA cytoskeleton regulatory RNA targeting microRNA-1246 on cell damage in Parkinson’s disease models

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