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

蔡梅芝; 马有权

doi:10.16098/j.issn.0529-1356.2022.05.005

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解剖学报 ›› 2022, Vol. 53 ›› Issue (5) : 571-577. DOI: 10.16098/j.issn.0529-1356.2022.05.005

神经生物学

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

蔡梅芝^1*卢宝全¹ 吴秀玲^１石佳¹ 马有权²

作者信息 +

Effect of long non-coding RNA nuclear-enriched abundant transcript on hypoxi-reoxygenation rat astrocyte injury by targeting microRNA-761

CAI Mei-zhi^1* LU Bao-quan¹ WU Xiu-ling¹ SHI Jia¹ MA You-quan²

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2022.05.005

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 https://doi.org/10.16098/j.issn.0529-1356.2022.05.005

中图分类号： R364.4

参考文献

［1］Liu H, Wu X, Luo J, et al. Pterostilbene attenuates astrocytic inflammation and neuronal oxidative injury after ischemia-reperfusion by inhibiting NF-κB phosphorylation ［J］. Front Immunol, 2019, 10(1): 2408.

［2］Schmitz SU, Grote P, Herrmann BG. Mechanisms of long noncoding RNA function in development and disease［J］. Cell Mol Life Sci, 2016, 73(13): 2491-2509.

［3］Zhou ZW, Ren X, Zheng LJ, et al. LncRNA NEAT1 ameliorate ischemic stroke via promoting Mfn2 expression through binding to Nova and activates Sirt3［J］. Metab Brain Dis, 2022, 37(3):653-664.

［4］Xie SP, Zhou F, Li J, et al. NEAT1 regulates MPP+-induced neuronal injury by targeting miR-124 in neuroblastoma cells［J］. Neurosci Lett, 2019, 708(1): 134340-134350.

［5］Long B, Wang K, Li N, et al. MiR761 regulates the mitochondrial network by targeting mitochondrial fission factor［J］. Free Radic Biol Med, 2013, 65(1): 371-379.

［6］Xu DCh, Fang LL, Chen H, et al. Effects of salidroside on hypoxia-reoxygenation astrocyte damage［J］. Chinese Journal of Immunology, 2019, 35(11): 1325-1330. (in chinese)

许德超, 方丽丽, 陈辉, 等. 红景天苷对缺氧复氧星形胶质细胞细胞损伤的影响［J］. 中国免疫学杂志, 2019, 35(11): 1325-1330.

［7］Wang SM, Liu GQ, Xian HB, et al. LncRNA NEAT1 alleviates sepsis-induced myocardial injury by regulating the TLR2/NF-κB signaling pathway［J］. Eur Rev Med Pharmacol Sci, 2019, 23(11): 4898-4907.

［8］Zhang M, Wang X, Yao J, et al. Long non-coding RNA NEAT1 inhibits oxidative stress-induced vascular endothelial cell injury by activating the miR-181d-5p/CDKN3 axis［J］. Artif Cells Nanomed Biotechnol, 2019, 47(1): 3129-3137.

［9］Chen Y, Qiu J, Chen B, et al. Long non-coding RNA NEAT1 plays an important role in sepsis-induced acute kidney injury by targeting miR-204 and modulating the NF-κB pathway［J］. Int Immunopharmacol, 2018, 59(1): 252-260.

［10］Ren L, Chen S, Liu W, et al. Downregulation of long non-coding RNA nuclear enriched abundant transcript 1 promotes cell proliferation and inhibits cell apoptosis by targeting miR-193a in myocardial ischemia/reperfusion injury ［J］. BMC Cardiovasc Disord, 2019, 19(1): 192.

［11］Du XJ, Wei J, Tian D, et al. NEAT1 promotes myocardial ischemia-reperfusion injury via activating the MAPK signaling pathway［J］. J Cell Physiol, 2019, 234(10): 18773-18780.

［12］Han D, Zhou Y. YY1-induced upregulation of lncRNA NEAT1 contributes to OGD/R injury-induced inflammatory response in cerebral microglial cells via Wnt/β-catenin signaling pathway ［J］. In Vitro Cell Dev Biol Anim, 2019, 55(7): 501-511.

［13］Shen S, Ma L, Shao F, et al. Long non-coding RNA (lncRNA) NEAT1 aggravates cerebral ischemia-reperfusion injury by suppressing the inhibitory effect of miR-214 on PTEN［J］. Med Sci Monit, 2020, 26(1): e924781.

［14］Zhou S, Zhang D, Guo J, et al. Deficiency of NEAT1 prevented MPP+-induced inflammatory response, oxidative stress and apoptosis in dopaminergic SK-N-SH neuroblastoma cells via miR-1277-5p/ARHGAP26 axis［J］. Brain Res, 2020, 15(1): 147156-147166.

［15］Yang YCh, Wang RF, Chen XJ, et al. Expression changes of Bcl-2 and Bax after cerebral ischemia-reperfusion injury in hyperlipidemia rats and the effect of scutellarin［J］. Acta Anatomy Sinica, 2019, 50(4): 431-437. (in Chinese)

杨迎春, 王瑞芳, 陈新骥, 等. 高血脂大鼠脑缺血再灌注损伤后Bcl-2和Bax的表达变化及灯盏乙素的影响［J］. 解剖学报, 2019, 50(4): 431-437.

［16］Jiang X, Li D, Shen W, et al. LncRNA NEAT1 promotes hypoxia-induced renal tubular epithelial apoptosis through downregulating miR-27a-3p［J］. J Cell Biochem, 2019, 120(9): 16273-16282.

［17］Wei JL, Wu CJ, Chen JJ, et al. LncRNA NEAT1 promotes the progression of sepsis-induced myocardial cell injury by sponging miR-144-3p［J］. Eur Rev Med Pharmacol Sci, 2020, 24(2): 851-861.

［18］Xiang X, Zheng L, Li X. Silencing of lncRNA ZFAS1 protects against hypoxia/reoxygenation-induced injury in HL-1 cells through targeting the miR-761/CDIP1 axis［J］. J Cardiovasc Pharmacol, 2020, 76(5): 564-573.