Krüppel样因子4抑制高糖刺激MES-13细胞炎症的机制  

Research of Krüppel-like factor 4 inhibit inflammation in MES-13 cells stimulated by high glucose

作　　者：黄志华 袁琼婧[1] 唐文彬[1] 于平平[1] 方茜 杨婕[1] 曾湘丽 张海侠 杰玛 肖湘成[1] HUANG Zhihua;YUAN Qiongjing;TANG Wenbin;YU Pingping;FANG Qian;YANG Jie;ZENG Xiangli;ZHANG Haixia;JIE Ma;XIAO Xiangcheng(Department of Nephrology,Xiangya Hospital Center South University,Changsha 410008,China;Department of Nephrology,The Third Hospital of Changsha,Changsha 410015,China)

机构地区：[1]中南大学湘雅医院肾内科,长沙410008 [2]长沙市第三医院肾内科

出　　处：《肾脏病与透析肾移植杂志》2018年第4期336-341,343,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：湖南省自然科学基金面上项目(2016JJ6161)

摘　　要：目的:观察Krüppel样因子4(KLF4)在小鼠肾小球系膜细胞(MES-13细胞)中的表达。观察甲基化抑制剂5-氮杂胞苷(5-azacytidine,5-Az)对高糖刺激的MES-13细胞中KLF4、IL-6、MCP-1表达的影响。方法:常规培养MES-13细胞,经高糖(30 mmol/L)处理不同时间后,检测KLF4蛋白表达水平。将细胞分为低糖组、高渗组、高糖组、高糖+不同浓度5-Az组,检测KLF4、IL-6、MCP-1 mRNA和蛋白表达水平。慢病毒质粒转染细胞后,高糖处理0h、48h,检测KLF4、IL-6、MCP-1 mRNA表达。结果:高糖处理MES-13细胞后,KLF4蛋白表达于24 h开始下调,于48h时下调明显。高糖处理细胞48h后,KLF4 mRNA和蛋白表达明显下调,MCP-1、IL-6 mRNA和蛋白表达水平明显上调(P<0.01),加用5-Az(5μmol/L)干预后,KLF4 mRNA和蛋白表达水平明显上调,而IL-6、MCP-1mRNA和蛋白表达均较干预前下调。慢病毒质粒转染细胞后,KLF4 mRNA表达明显增加,IL-6、MCP-1 mRNA表达无明显变化。高糖处理转染细胞48h后,Lv-NC(hi)组与Lv-NC组相比,KLF4 mRNA表达水平明显下调,IL-6、MCP-1 mRNA表达水平明显上调。Lv-KLF4 (hi)组与Lv-NC (hi)组相比,KLF4 mRNA表达水平明显上调,IL-6、MCP-1 mRNA表达水平明显下调。结论:在MES-13细胞中存在KLF4表达,且高糖处理可使KLF4表达下调,IL-6、MCP-1表达上调。高糖引起的MES-13细胞炎症与KLF4低表达有关,恢复KLF4表达,可减轻细胞炎症,推测其机制部分与DNA甲基化相关。Objective：To observe the expression of Krtippel-like factor 4 （KLF4） in MES-13 cells and to detect the expression of KLF4, IL-6, MCP-lin MES-13 ceils stimulated by high glucose. Methodology： MES-13 ceils were stimulated by high glucose （30 mmoL/L） for different time. The expression of KLF4 protein was detected by Western blot respeetively.MES-13 cells were divided into four groups： normal glucose, hypertonic group, high glucose and high glucose with 5-Az. The expressions of KLF4, IL-6, MCP-1 mRNA and protein were detected by RT-PCR and Western blot separately. MES-13 cells were divided into two groups：Lv-NC glucose and Lv-KLF4 group.The expressions of KLF4,IL-6, MCP-1 mRNA were detected by RT-PCR respectively after stimulating by high glucose for 0 hour and 48 hours. Results： In MES-13 cells, the expression of KLF4 protein was decreased after stimulating by high glucose for 24 hours, and it was significantly decreased at 48 hours. The expressions of KLF4 protein and mRNA were significantly decreased in MES-13 cells stimulated by high glucose for 48 hours, while the expressions of IL-6, MCP-1 protein and mRNA were increased obviously.After treatment with 5-Az （5 p, mol/L） , the expressions of KLF4 protein and mRNA were up-regulated in MES-13 cells,while the expressions of IL-6, MCP-1 protein and mRNA were down-regulated distinctly. After lentivirus plasmid transfected into MES-13 cells, the expression level of KLF4 mRNA was significantly increased, while IL-6 and MCP-1 mRNA expressions didn＇t change obviously.After stimulating MES-13 cells by high glucose for 48 hours ,the expression of KLF4 mRNA was significantly decreased in lv-nc （hi） group compared with lv-nc group, and the expression of IL-6 and MCP-1 mRNA were distinctly increased. Compared with lv-nc （ hi ） group, the expression level of KLF4 mRNA was significantly up-regulated in LV-KLF4 （hi） group, while the expression of IL-6 and MCP-1 mRNA were down-regulated obviously, Conelusion：KLF4 is expressed in MES-13 cells.H

关 键 词：糖尿病肾病 Krüppel样因子4 炎症反应 5-氮杂胞苷 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]