摘要
目的探讨长链非编码RNA(lncRNA)Loc102553943在大鼠缺氧神经元细胞中的表达及其对神经元细胞凋亡的影响。方法原代神经元细胞分为缺氧组(糖氧剥夺培养)、对照组(正常培养)和缺氧+lncRNA Loc102553943小干扰RNA(siRNA)组(缺氧后沉默lncRNA Loc102553943)。实时定量反转录聚合酶链反应(RT-qPCR)检测缺氧组和对照组神经元细胞中lncRNA Loc102553943的表达水平,流式细胞仪检测神经元细胞凋亡。缺氧神经元细胞经沉默表达lncRNA Loc10255394后,流式细胞技术检测细胞凋亡,蛋白质印迹法(Western blot)检测凋亡相关蛋白[B细胞淋巴瘤/白血病-2相关X蛋白(bax)、B细胞淋巴瘤/白血病-2(bcl-2)]。结果缺氧组lncRNA Loc102553943表达(4.37±0.32)较对照组(1.12±0.09)显著升高(P<0.05),且凋亡率明显增加[(4.26±0.08)%比(2.13±0.06)%,P<0.05];缺氧神经元细胞经沉默表达lncRNA Loc10255394后较缺氧组神经细胞凋亡率下降[(3.23±0.04)%比(5.36±0.08)%,P<0.05],而bcl-2蛋白表达上调[(0.92±0.16)%比(0.57±0.07)%,P<0.05]、bax蛋白表达下调[(1.08±0.08)%比(1.56±0.02)%,P<0.05]。结论大鼠缺氧神经元细胞中lncRNA Loc102553943表达上调,lncRNA Loc102553943上调促进缺氧后神经元细胞凋亡;而下调lncRNA Loc102553943后,神经元细胞凋亡随之减少。
Objective To explore the long non-coding RNA(lncRNA)102553943 expression in rat hypoxic neuronal cells and its effect on neuronal apoptosis.Methods Primary neuronal cells were divided glucose and oxygen deprivation group,normal culture group and glucose and oxygen deprivation+lncRNA Loc102553943 small interfering RNA(siRNA)group.The expression of lncRNA Loc102553943 was measured in glucose and oxygen deprivation group and normal culture group.Cell apoptosis was determined by flow in the two groups.The apoptosis of hypoxic neuronal cells after silent expression of lncRNA Loc10255394 was measured by flow and apoptosis-associated proteins[B cell lymphoma/leukemia-2 associated X protein(bax),B cell lymphoma/leukemia-2(bcl-2)]was measured by Western blotting.Results The expression of lncRNA Loc102553943 in the hypoxia group(4.37±0.32)was significantly higher than that of the control group(1.12±0.09,P<0.05)and the apoptosis rate was also significantly higher[(4.26±0.08)%vs.(2.13±0.06)%,P<0.05];The apoptosis rate of hypoxic neuronal cells after silent expression of lncRNA Loc10255394 was lower than that of the hypoxic group[(3.23±0.04)%vs.(5.36±0.08)%,P<0.05],while the expression of bcl-2 protein was upregulated[(0.92±0.16)%vs.(0.57±0.07)%,P<0.05],and bax protein expression was down-regulated[(1.08±0.08)%vs.(1.56±0.02)%,P<0.05].Conclusion lncRNA Loc102553943 is upregulated in rat hypoxic neuronal cells and promotes apoptosis after hypoxia.
作者
陈丽
黄宽
梁伟东
王力峰
曾文
郭锐
Chen Li;Huang Kuan;Liang Weidong;Wang Lifeng;Zen Wen;Guo Rui(Department of Anesthesiology,the First Affiliated Hospital of Gannan Medical College,Ganzhou 341000,China;Gannan Medical College,Ganzhou 341000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第12期2432-2434,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(81260202)
心脑血管疾病防治教育部重点实验室赣南医学院科技项目(XN201917)。
