长链非编码RNA-N1LR在脑缺血再灌注血脑屏障损伤的作用机制研究  

Mechanism of lncRNA-N1LR in blood-brain barrier injury during cerebral ischemia reperfusion

作　　者：胡云 周礼鑫 童理 李鑫泰 杨剑文[1] Hu Yun;Zhou Lixin;Tong Li;Li Xintai;Yang Jianwen(Department of Neurology,Hunan Provincial People's Hospital The First Affiliated Hospital of Hunan Normal University,Changsha 410015,Hunan Province,China)

机构地区：[1]湖南省人民医院(湖南师范大学附属第一医院)神经内科,长沙410015

出　　处：《中华老年心脑血管病杂志》2024年第2期217-220,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：湖南省卫生健康委科技资助项目(202203072442);湖南省教育厅科技资助项目(20C1135)。

摘　　要：目的探讨长链非编码RNA(lncRNA)-N1LR在脑缺血再灌注损伤后血脑屏障的作用机制。方法原代小鼠脑微血管内皮细胞常规培养,经氧糖剥夺/复糖复氧(OGD/R)处理模拟脑缺血再灌注损伤,实验分对照组、OGD组、lncRNA-N1LR过表达组(OGD处理后转染质粒lncRNA-N1LR过表达)、lncRNA-N1LR沉默组(OGD处理后转染质粒lncRNA-N1LR沉默)。采用逆转录聚合酶链反应检测各组中lncRNA-N1LR mRNA、紧密连接蛋白5(claudin-5)及闭合蛋白(occludin)mRNA表达水平;异硫氰酸荧光素-葡聚糖(FITC-dextran)渗透法检测血脑屏障通透性;免疫蛋白印迹法检测claudin-5、occludin蛋白表达。结果与对照组比较,OGD组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平下降(0.31±0.01 vs 1.00±0.10,0.42±0.03 vs 1.01±0.13,0.38±0.03 vs 1.00±0.15,P<0.05),血脑屏障FITC-dextran通透性明显升高(58.79±3.04 vs 8.87±0.63,P<0.01)。与OGD组比较,lncRNA-N1LR过表达组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平升高(0.67±0.07 vs 0.31±0.01,0.92±0.02 vs 0.42±0.03,0.70±0.08 vs 0.38±0.03,P<0.05),血脑屏障FITC-dextran通透性降低(41.57±2.43 vs 58.79±3.04,P<0.05)。与OGD组比较,lncRNA-N1LR沉默组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平降低(0.21±0.02 vs 0.31±0.01,0.31±0.03 vs 0.42±0.03,0.22±0.02 vs 0.38±0.03,P<0.05),血脑屏障FITC-dextran通透性升高(72.34±1.43 vs 58.79±3.04,P<0.05)。结论LncRNA-N1LR上调可能通过降低血脑屏障通透性发挥神经保护作用。Objective To investigate the action mechanism of long non-coding RNA(lncRNA)-N1LR on blood-brain barrier(BBB)after cerebral ischemia-reperfusion(I/R)injury.Methods Primary rat brain microvascular endothelial cells(BMECs)were cultured and treated with OGD/R to simulate cerebral I/R injury.The experiment was divided into normal control group,lncRNA-N1LR OGD group,overexpression group(lncRNA-N1LR overexpression after OGD treatment)and silence group(lncRNA-N1LR silence after OGD treatment).The mRNA levels of lncRNA-N1LR,claudin-5 and occludin in each group were detected by RT-qPCR.The BBB permeability was detected by FITC-dextran infiltration assay.The expression of claudin-5 and occludin were detected by Western blotting.Results The mRNA levels of lncRNA-N1LR,occludin and claudin-5 were significantly decreased(0.31±0.01 vs 1.00±0.10,0.42±0.03 vs 1.01±0.13,0.38±0.03 vs 1.00±0.15,P<0.05),and the BBB permeability was significantly increased(58.79±3.04 vs 8.87±0.63,P<0.05)in the OGD group than the control group.The lncRNA-N1LR overexpression group increased the mRNA expression of lncRNA-N1LR,occludin and claudin-5(0.67±0.07 vs 0.31±0.01,0.92±0.02 vs 0.42±0.03,0.70±0.08 vs 0.38±0.03,P<0.05),and decreased the BBB permeability(41.57±2.43 vs 58.79±3.04,P<0.05)than the OGD group.lncRNA-N1LR silence resulted in lower mRNA levels of lncRNA-N1LR,occludin and claudin-5(0.21±0.02 vs 0.31±0.01,0.31±0.03 vs 0.42±0.03,0.22±0.02 vs 0.38±0.03,P<0.05),and enhanced BBB permeability(72.34±1.43 vs 58.79±3.04,P<0.05)when compared with the OGD group.Conclusion Up-regulation of lncRNA-N1LR may play a neuroprotective role by reducing BBB permeability.

关 键 词：RNA 未翻译 再灌注损伤 血脑屏障 神经保护 MICE 模型 动物 长链非编码RNA-N1LR 

分 类 号：R743.3[医药卫生—神经病学与精神病学]