LINC01116靶向miR-203加剧氧糖剥夺/复氧诱导下人海马星形胶质细胞损伤  

LINC01116 aggravates oxygen-glucose deprivation/reoxygenation-induced injury in human hippocampal astrocytes by targeting miR-203

作　　者：黄正义 赵增霞 符秋红 刘淑云 陈协辉 HUANG Zhengyi;ZHAO Zengxia;FU Qiuhong(Department of Geriatrics,Shenzhen Longhua District Central Hospital,Shenzhen 518110,China)

机构地区：[1]深圳市龙华区中心医院老年医学科,广东深圳518110 [2]深圳市龙华区中心医院神经内科,广东深圳518110 [3]深圳市龙华区中心医院急诊科,广东深圳518110

出　　处：《中风与神经疾病杂志》2022年第10期887-892,共6页Journal of Apoplexy and Nervous Diseases

基　　金：深圳市龙华区医疗卫生机构区级科研项目(No.2021002)。

摘　　要：目的明确长基因间非编码RNA 1116(LINC01116)在急性缺血性脑卒中(AIS)海马星形胶质细胞中的作用及分子机制。方法利用实时荧光定量PCR检测131例AIS患者溶栓前后血清中LINC01116与miR-203表达。采用双荧光素酶报告基因与RNA结合蛋白免疫沉淀试验检测LINC01116对miR-203的调控作用。以人海马星形胶质细胞(hHA)建立氧糖剥夺/复氧(OGD/R)模型并给予LINC01116干扰与LINC01116联合miR-203干扰处理,采用CCK-8、TUNEL与蛋白质印迹、活性氧(ROS)试验、乳酸脱氢酶(LDH)试验及酶联免疫吸附试验分别检测细胞增殖、细胞凋亡、ROS生成、LDH活性及炎症因子的变化。结果溶栓后血清中LINC01116与miR-203的表达分别高于与低于溶栓前,且LINC01116与miR-203表达呈负相关(P<0.05)。LINC01116海绵吸附miR-203并抑制其表达(P<0.05)。LINC01116干扰减轻OGD/R诱导下hHA细胞损伤,表现为细胞增殖能力提高,细胞凋亡率下降,激活型caspase-3与Bax蛋白表达降低而Bcl-2蛋白表达升高,ROS生成减少,LDH活性下降,TNF-α、IL-1β及IL-6浓度下调而IL-4、IL-10及IL-13浓度上调(P<0.05)。miR-203干扰逆转LINC01116干扰对OGD/R诱导下hHA细胞损伤的保护作用(P<0.05)。结论LINC01116靶向miR-203促进OGD/R诱导下hHA细胞损伤,表明LINC01116/miR-203通路可能是AIS的潜在治疗靶点。Objective To investigate the role and molecular mechanism of long intergenic non-coding RNA 1116(LINC01116)in hippocampal astrocytes of acute ischemic stroke(AIS).Methods The expressions of LINC01116 and miR-203 in serum of 131 AIS patients before and after thrombolysis were detected by real-time fluorescence quantitative PCR.The regulatory effect of LINC01116 on miR-203 was detected by dual-luciferase reporter gene and RNA-binding protein immunoprecipitation assay.Human hippocampal astrocytes(hHA)were applied to establish an oxygen-glucose deprivation/reoxygenation(OGD/R)model and were treated with LINC01116 interference and LINC01116 combined with miR-203 interference.The changes of cell proliferation,cell apoptosis,production of reactive oxygen species(ROS),activity of lactate dehydrogenase(LDH),and inflammatory factors were detected by CCK-8,TUNEL and Western blotting,ROS assay,LDH assay,and enzyme-linked immunosorbent assay,respectively.Results The expressions of LINC01116 and miR-203 in serum after thrombolysis were higher and lower than those before thrombolysis,respectively,and the expressions of LINC01116 and miR-203 were negatively correlated(P<0.05).LINC01116 inhibited miR-203 expression by sponge of miR-203(P<0.05).LINC01116 interference alleviated the OGD/R-induced injury of hHA cells,which manifested as elevated cell proliferation ability,decreased cell apoptosis rate,decreased protein expressions of cleaved caspase-3 and Bax but raised protein expression of Bcl-2,reduced ROS production,decreased LDH activity,and downregulated TNF-α,IL-1βand IL-6 concentrations but upregulated IL-4,IL-10 and IL-13 concentrations(P<0.05).miR-203 interference reversed the protective effect of LINC01116 interference on the OGD/R-induced injury of hHA cells(P<0.05).Conclusions LINC01116 promotes the OGD/R-induced injury of hHA cells by targeting miR-203,suggesting that the LINC01116/miR-203 pathway might be a potential therapeutic target for AIS.

关 键 词：长基因间非编码RNA 1116 急性缺血性脑卒中 人海马星形胶质细胞 氧糖剥夺/复氧 治疗靶点 

分 类 号：R743.3[医药卫生—神经病学与精神病学]