长链非编码RNA在内皮细胞损伤模型中的表达调控研究  

Study on the Expression Regulation of Long Non-coding RNA in Endothelial Cell Injury Model

作　　者：张蕊 耿佳 王雪梅 ZHANG Rui;GENG Jia;WANG Xue-mei(Teaching and Research Office of Hygiene,Department of Public Health,Xi'an Medical University,Xi'an 710021,Shaanxi,China;the Key Laboratory of Animal Disease Model Research,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China)

机构地区：[1]西安医学院公共卫生学院卫生学教研室,陕西西安710021 [2]新疆医科大学第一附属医院动物疾病模型研究重点实验室,新疆乌鲁木齐830054

出　　处：《医学信息》2020年第22期71-74,95,共5页Journal of Medical Information

基　　金：国家自然科学基金地区基金项目(编号:81760083);西安医学院大学生创新创业训练计划项目(编号:省级;校级<121519049>);陕西省教育厅重点实验室项目(编号:20JS137)。

摘　　要：目的研究不同应激条件下的内皮细胞(HUVECs)损伤模型中lncRNAs的表达变化。方法将HUVECs分为正常对照组、H2O2干预组、LPS诱导组及ox-LDL干预组,正常对照组细胞常规培养,H2O2干预组采用200μmol/L浓度的H2O2孵育12 h,LPS诱导组采用100 ng/ml LPS诱导6 h,ox-LDL干预组使用浓度为50μg/ml ox-LDL的培养液诱导内皮细胞损伤48 h。建立模型后分别检测心梗相关lncRNA H19、MAIT、MALAT1、ANRIL在脐静脉内皮细胞中的m RNA表达;实时荧光定量法测定HUVECs中lncRNAs的表达,以及炎性因子VCAM-1、MCP-1和抗炎因子eNOS、IL-10、Arginine的m RNA表达;ELISA法检测培养上清液中促炎因子及抗炎因子的表达。结果与对照组相比,ox-LDL干预组lncRNA H19、MAIT、ANRIL的表达均上调,差异有统计学意义(P<0.05);炎性因子MCP-1和VCAM-1的m RNA表达水平均上调(P<0.05),eNOS的m RNA表达下调(P<0.05),细胞上清液中的MCP-1、VCAM-1的表达增加(P<0.05)。ox-LDL干预组的HUVECs细胞上清液中的eNOS活性低于空白对照组,差异有统计学意义(P<0.05)。结论 lncRNA H19、MIAT、MALAT1、ANRIL参与调控了内皮细胞的脂质代谢。Objective To study the expression changes of lncRNAs in endothelial cell(HUVECs)injury models under different stress conditions.Methods HUVECs were divided into normal control group,H2O2 intervention group,LPS induction group and ox-LDL intervention group.Normal control group cells were cultured routinely.The H2O2 intervention group was incubated with 200μmol/L H2O2 for 12 h,and the LPS induction group was 100 ng/ml LPS was induced for 6 h,and the ox-LDL intervention group used a culture medium with a concentration of 50μg/ml ox-LDL to induce endothelial cell damage for 48 h.After the model was established,the mRNA expression of myocardial infarction-related lncRNA H19,MAIT,MALAT1,and ANRIL in umbilical vein endothelial cells were detected;real-time fluorescence quantitative method was used to determine the expression of lncRNAs in HUVECs,as well as the inflammatory factors VCAM-1,MCP-1 And anti-inflammatory factors eNOS,IL-10,Arginine mRNA expression;ELISA method to detect the expression of pro-inflammatory factors and anti-inflammatory factors in the culture supernatant.Results Compared with the control group,the expressions of lncRNA H19,MAIT and ANRIL in the ox-LDL intervention group were all up-regulated,the difference was statistically significant(P<0.05);the mRNA expression levels of inflammatory factors MCP-1 and VCAM-1 were both up-regulated(P<0.05),eNOS mRNA expression was down-regulated(P<0.05),and the expression of MCP-1 and VCAM-1 in the cell supernatant increased(P<0.05).The eNOS activity in the HUVECs cell supernatant of the ox-LDL intervention group was lower than that of the blank control group,the difference was significant(P<0.05).Conclusion lncRNA H19,MIAT,MALAT1,ANRIL participate in the regulation of lipid metabolism of endothelial cells.

关 键 词：长链非编码RNA 动脉粥样硬化 人脐静脉内皮细胞 氧化型低密度脂蛋白 

分 类 号：R541.4[医药卫生—心血管疾病]