长链非编码RNA 核富集转录本1通过靶向微小RNA-761对缺氧/复氧大鼠星形胶质细胞损伤的影响

doi:10.16098/j.issn.0529-1356.2022.05.005

摘要/Abstract

摘要：

目的探讨长链非编码RNA（lncRNA）核富集转录本1（NEAT1）对缺氧/复氧（H/R）胶质细胞损伤的影响，及其机制是否与调控微小RNA-761（miR-761）有关。方法构建大鼠皮质星形胶质细胞H/R损伤模型。分为对照组、H/R组、H/R+小干扰RNA阴性对照（si-NC）组、H/R+si-NEAT1组、H/R+miR-NC组、H/R+miR-761组、H/R+si-NEAT1+ miRNA抑制物（anti-miR-NC）组、H/R+si-NEAT1+anti-miR-761组。Real-time PCR分析NEAT1和miR-761的mRNA表达。流式细胞术分析细胞凋亡。试剂盒检测细胞中丙二醛（MDA）含量以及超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。双荧光素酶报告基因实验和Real-time PCR分析NEAT1和miR-761靶向关系。实验重复3次。结果与对照组比较，H/R组细胞凋亡率和MDA含量显著升高，SOD和CAT活性显著降低，NEAT1表达显著升高，miR-761表达显著降低（P<0.05）。与H/R+si-NC组比较，H/R+si-NEAT1组细胞凋亡率和MDA含量显著降低，SOD和CAT活性显著升高（P<0.05）。与H/R+miR-NC组比较，H/R+miR-761组细胞凋亡率和MDA含量显著降低，SOD和CAT活性显著升高（P<0.05）。MiR-761是NEAT1的靶基因，NEAT1负调控miR-761表达。与 H/R+si-NEAT1+anti-miR-NC 组比较，H/R+si-NEAT1+anti-miR-761组细胞凋亡率、MDA含量显著升高，SOD和CAT活性显著降低（P<0.05）。结论干扰NEAT1表达通过上调miR-761对H/R损伤的星形胶质细胞具有保护作用。

关键词: 长链非编码RNA, 核富集转录本1, 星形胶质细胞, 缺氧/复氧, 微小RNA-761, 实时定量聚合酶链反应, 流式细胞术

Abstract:

Objective To investigate the effect of long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) on hypoxia-reoxygenation(H/R) glial astrocyte injury, and to explore whether the mechanism was related to the regulation of micro RNA (miR)-761. Methods Rat cortical astrocytes were cultured to construct a H/R injury model. Astrocytes were divided into control group, model group, model+ small interfering RNA negative control (si-NC) group, model+si-NEAT1 group, model+miR-NC group, model+miR-761 group, model+si-NEAT1+anti-miR-NC group, model+si-NEAT1+anti-miR-761 group. Expression of NEAT1 and miR-761 were detected by Real-time PCR. The experiment was repeated 3 times. The content of malonaldehyde (MDA), and the activity of superoxide dismutase (SOD) and catalase (CAT) were detected by kits. Dual luciferase reporter experiment and Real-time PCR were used to analyze the targeting relationship between NEAT1 and miR-761. The experiment was repeated 3 times. Results Compared with the control group, the cell apoptosis rate and MDA content of the model group increased significantly, SOD and CAT activities decreased significantly, NEAT1 expression increased significantly, and miR-761 expression decreased significantly (P<0.05). Compared with the model+si-NC group, the apoptosis rate and MDA content of the model+si-NEAT1 group reduced significantly, and SOD and CAT activities increased significantly (P<0.05). Compared with the model+miR-NC group, the apoptosis rate and MDA content of the model+miR-761 group reduced significantly, and SOD and CAT activities increased significantly (P<0.05). MiR-761 was the target gene of NEAT1, and NEAT1 negatively regulated miR-761 expression. Compared with the model+si-NEAT1+anti-miR-NC group, the apoptosis rate and MDA content of the model+si-NEAT1+anti-miR-761 group increased significantly, and SOD and CAT activities decreased significantly (P<0.05). Conclusion Interference with NEAT1 expression can protect astrocytes from H/R injury by up-regulating miR-761.

Key words: Long non-coding RNA, Nuclear-enriched abundant transcript 1, Astrocyte, Hypoxia-reoxygenation, MicroRNA-761, Real-time PCR, Flow cytometry

中图分类号:

R364.4

蔡梅芝卢宝全吴秀玲石佳马有权. 长链非编码RNA 核富集转录本1通过靶向微小RNA-761对缺氧/复氧大鼠星形胶质细胞损伤的影响[J]. 解剖学报, 2022, 53(5): 571-577.

CAI Mei-zhi LU Bao-quan WU Xiu-ling SHI Jia MA You-quan . Effect of long non-coding RNA nuclear-enriched abundant transcript on hypoxi-reoxygenation rat astrocyte injury by targeting microRNA-761[J]. Acta Anatomica Sinica, 2022, 53(5): 571-577.

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