EZH2基因表达对ox-LDL诱导内皮细胞炎症反应的影响  

Effect of EZH2 gene expression on inflammatory response of endothelial cells induced by ox-LDL

作　　者：李扬 林飞[1,2,3] 陈志刚 常峥峥[1] 赵奕霖 李东旭 李雪芳 孙四玉 赵国安 LI Yang;LIN Fei;CHENG Zhigang;CHANG Zhengzheng;ZHAO Yilin;LI Dongxu;LI Xuefang;SUN Siyu;ZHAO Guoan(Life Science Research Center of the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,China;不详)

机构地区：[1]新乡医学院第一附属医院生命科学研究中心,河南新乡453100 [2]河南省心血管损伤与修复国际联合实验室 [3]河南省心脏生物医学工程研究中心

出　　处：《山东医药》2021年第23期10-13,32,共5页Shandong Medical Journal

基　　金：河南省高等学校重点科研项目(19A360032);河南省医学科技攻关计划(LHGJ20190441)。

摘　　要：目的观察Zeste同源复合物2(EZH2)基因表达对氧化低密度脂蛋白(ox-LDL)诱导内皮细胞炎症反应的影响。方法培养人脐静脉内皮细胞(HUVEC),加入不同浓度(0、25、50、100μg/mL)ox-LDL作用12、24 h,采用ELISA法检测细胞上清白细胞介素6(IL-6)水平;结果显示,100μg/mL ox-LDL作用后上清IL-6水平升高最明显(P均<0.05),因0μg/mL ox-LDL作用24 h后上清IL-6水平升高,故选择100μg/mL ox-LDL作用12 h作为后续实验条件。将HUVEC分为两部分,一部分分为实验A组(EZH2腺病毒+100μg/mL ox-LDL)、阳性对照A组(对照腺病毒+100μg/mL ox-LDL)、阴性对照A组(100μg/mL ox-LDL)及空白A组(未处理),另一部分分实验B组(EZH2特异性抑制剂GSK126+100μg/mL ox-LDL)、阳性对照B组(1%DMSO+100μg/mL ox-LDL)、阴性对照B组(100μg/mL oxLDL)及空白B组(未处理),均作用12 h。采用Western blotting法检测细胞EZH2蛋白表达,ELISA法检测上清IL-6水平,Real-time PCR法检测细胞EZH2及IL-6、肿瘤坏死因子α(TNF-α)、一氧化氮合酶(e-NOS)、IL-8、C反应蛋白(CRP)mRNA表达。结果与空白A组比较,实验A组细胞EZH2 mRNA、蛋白表达明显升高(P均<0.05),阳性对照A组、阴性对照A组细胞EZH2 mRNA、蛋白表达无明显变化(P均>0.05)。与空白A组比较,实验A组、阳性对照A组、阴性对照A组上清IL-6水平及细胞TNF-α、e-NOS、IL-8 mRNA表达均升高,实验A组升高更明显(P均<0.05)。与空白B组比较,实验B组细胞EZH2 mRNA、蛋白表达明显降低(P均<0.05),阳性对照B组、阴性对照B组细胞EZH2 mRNA、蛋白表达无明显变化(P均>0.05)。与空白B组比较,实验B组、阳性对照B组、阴性对照B组上清IL-6水平及细胞TNF-α、e-NOS、IL-8 mRNA表达均升高,阳性对照B组、阴性对照B组升高更明显(P均<0.05)。各组细胞CRP mRNA表达比较均无统计学差异(P均>0.05)。结论 EZH2过表达可通过靶向调节炎症因子TNF-α、eNOS、IL-8、IL-6而促进ox-LDL诱导内皮细胞炎症反应,抑制EZH2表Objective To explore the influence of Zeste homologous complex 2(EZH2)gene expression on endothe-lial cell inflammation induced by oxidized low-density lipoprotein(ox-LDL).Methods Human umbilical vein endotheli-al cells(HUVEC)were cultured and induced by ox-LDL at different concentrations(0,25,50,100μg/mL)for 12 and 24 h.The level of interleukin 6(IL-6)in the supernatant was detected by ELISA.The results showed that at the same time,the level of IL-6 in the supernatant of HUVEC treated with 25,50,and100μmol/L ox-LDL was significantly higher than that of HUVEC treated with 0μmol/L ox-LDL,and the level of IL-6 in the supernatant increased significantly after treatment with 100μg/mL ox-LDL.Because the level of IL-6 in the supernatant increased after 24 h treatment with 0μg/mL ox-LDL,100μg/mL ox-LDL for 12 h was chosen as the follow-up experimental condition.HUVEC were randomly divided into two parts:the first part was divided into the experimental group A(EZH2 adenovirus+100μg/mL ox-LDL),positive control group A(control adenovirus+100μg/mL ox-LDL),negative control group A(100μg/mL ox-LDL),and blank group A(untreated).The other part was divided into the experimental group B(EZH2 specific inhibitor GSK126+100μg/mL ox-LDL),positive control group B(1%DMSO+100μg/mL ox-LDL),negative control group B(100μg/mL ox-LDL)and blank group B(untreated)for 12 h.Western blotting was applied to detect the expression of EZH2 protein,ELISA was applied to detect the level of IL-6 in the supernatant,and PCR was applied to detect and the expression levels of EZH2 and IL-6,TNF-α,e-NOS,IL-8 and CRP mRNA.Results Compared with the blank group A,the expression levels of EZH2 mRNA and protein in the experimental group A increased,while there were no significant changes in the ex-pression levels of EZH2 mRNA and protein in the positive control group A and negative control group A(all P>0.05).Compared with the blank group A,the level of IL-6 in the supernatant and the expression of TNF-α,e-NOS,IL-8 and IL-6 mRNA increased in the expe

关 键 词：动脉粥样硬化 内皮细胞 炎症 EZH2 OX-LDL TNF-α e-NOS IL-8 IL-6 CRP 

分 类 号：R543.5[医药卫生—心血管疾病]