lncRNA H19和lncRNA GAS5在急性缺血性脑卒中的表达及临床意义

Expression and clinical significance of lncRNA H19 and lncRNA GAS5 in acute ischemic stroke

目的本研究旨在检测长链非编码RNA H19和生长阻滞特异转录物5(GAS5)在急性缺血性脑卒中(IS)患者单个核细胞(PBMC)、大脑中动脉闭塞模型(MCAO)大鼠脑皮质组织和氧糖剥夺再灌注(OGD/R)模型PC12细胞的表达水平,探讨其作为IS分子诊断指标的可行性。方法收集23例IS组患者和33例健康对照外周血,提取PBMC,采用实时定量PCR(qPCR)检测PBMC中lncRNA H19和GAS5的表达水平。进一步构建大鼠MCAO模型和PC12细胞OGD/R模型,qPCR分别检测lncRNA H19和GAS5在大鼠脑皮质和PC12细胞中的表达。结果 qPCR结果显示,IS组患者PBMC的lncRNA H19表达水平显著高于对照组,差异有统计学意义(P<0.05)。IS组患者PBMC中的lncRNA GAS5表达水平显著高于对照组,差异有统计学意义(P<0.05)。和假手术组相比,MCAO模型大鼠缺血脑皮质lncRNA H19和lncRNA GAS5表达水平显著升高,差异有统计学意义(P<0.05)。细胞实验结果表明,PC12细胞在OGD/R后,lncRNA H19和lncRNA GAS5表达水平显著升高,差异有统计学意义(P<0.05)。结论lncRNA H19和lncRNA GAS5在IS表达升高,有望成为诊断IS的生物标志物。

Objective To examine the expression levels of long non-coding RNA(lncRNA)H19 and growth arrest-specific transcript 5(GAS5)in the peripheral blood mononuclear cells(PBMC)from acute ischemic stroke(IS)patients,cortex of rats with middle cerebral artery occlusion(MCAO)model,oxygen,and glucose deprivation/reoxygenation(OGD/R)-treated PC12 cells and further evaluate their potential as biomarkers for the diagnosis of IS.Methods Blood from 23 acute IS patients and 33 healthy controls were collected.PBMC were isolated by Ficoll lymphocytes separation medium.Quantitative real-time polymerase chain reaction(RT-qPCR)methods were used to detect the expression levels of lncRNA H19 and GAS5.Furthermore,rats were established by MCAO model and PC12 cells were subjected to OGD/R to simulate cerebral ischemia and reperfusion injury.The levels of lncRNA H19 and GAS5 were also assessed in cortex of MCAO rats and PC12 cells subjected to OGD/R using RT-qPCR.Results Compared with the healthy control,the expression levels of lncRNA H19 were significantly upregulated in IS patients(P<0.05).The expression levels of lnc RNA GAS5 were also significantly upregulated in IS patients(P<0.05).Compared with the sham control,the expression levels of lnc RNA H19 and lnc RNA GAS5 were significantly increased in the cortex from MCAO rats(P<0.05).The cell experiment results showed that the levels of lnc RNA H19 and lnc RNA GAS5 were increased significantly in OGD/R PC12 cells(P<0.05).The expressions of lnc RNA H19 and lnc RNA GAS5 in IS are elevated,and they are expected to be biomarkers for the diagnosis of IS.