miR-222调控PARP-1在高糖致人肾脏系膜细胞tPA/PAI-1紊乱中的作用  

Role of the regulation of PARP-1 by miR-222 in tPA/PAI-1 disturbance induced by high glucose in human mesangial cells

作　　者：朱恒梅[1] 祝胜郎[1] 陈结慧[1] 杨芸 李金艳[2] Zhu Hengmei;Zhu Shenglang;Chen Jiehui;Yang Yun;Li Jinyan(Department of Nephrology,Nanshan People＇s Hospital,Shenzhen 518067,China)

机构地区：[1]深圳市南山区人民医院肾内科,深圳518067 [2]深圳市南山区人民医院核医学科,深圳518067

出　　处：《新医学》2018年第7期502-506,共5页Journal of New Medicine

基　　金：深圳市科技计划项目(JCYJ20160429181842402);深圳市南山区科技计划项目(南科研卫2014054号)

摘　　要：目的探讨微小RNA-222(miR-222)调控多聚二磷酸腺苷核糖聚合酶-1(PARP-1)在高糖诱导人肾脏系膜细胞(HMC)组织型纤溶酶原激活物/纤溶酶原激活物抑制物-1(tPA/PAI-1)紊乱中的作用。方法体外培养HMC,实时定量PCR分别检测正常糖(5 mmol/L)及高糖(25 mmol/L)刺激下miR-222表达;另将体外培养的HMC分为正常糖组、25 mmol/L高糖组、高糖+miR-222 mimics阴性对照组、高糖+miR-222 mimics组4组,运用蛋白免疫印迹法检测PARP-1蛋白表达,ELISA法测定细胞上清液tPA、PAI-1的分泌蛋白。结果实时定量PCR显示,高糖组HMC在24 h时miR-222mRNA表达水平是正常糖组的2.537倍。高糖组HMC中PARP-1蛋白表达高于正常糖组(P<0.05),高糖+miR-222 mimics组HMC中PARP-1蛋白表达低于高糖组(P<0.05),而高糖+miR-222 mimics阴性对照组与高糖组比较差异无统计学意义(P>0.05)。与正常糖组比较,高糖组的tPA和tPA/PAI-1降低、PAI-1升高(P均<0.05);与高糖组比较,高糖+mimics组的tPA和tPA/PAI-1升高、PAI-1降低(P均<0.05)。高糖+miR-222 mimics阴性对照组与高糖组tPA、PAI-1及tPA/PAI-1比较差异均无统计学意义(P均>0.05)。结论高糖可增强HMC PARP-1及分泌型PAI-1的蛋白表达、减弱分泌型tPA的蛋白表达,下调tPA/PAI-1比值。miR-222可有效下调靶蛋白PARP-1的表达,并有效改善高糖诱导的tPA/PAI-1紊乱。Objective To investigate the role of the regulation of poly（ ADP-ribose） polymerase-1（ PARP-1） by microRNA-222（ miR-222） in the disturbance of tissue-type plasminogen activator/plasminogen activator inhibitor-1（ tPA/PAI-1） induced by high glucose in human mesangial cells（ HMCs）. Methods HMCs were cultured in vitro. Real-time PCR was adopted to detect the expression levels of miR-222 under the stimulation of normal glucose（ 5 mmol/L） and high glucose（ 25 mmol/L）. The HMCs cultured in vitro were divided into the normal glucose,25 mmol/L high glucose,high glucose ＋ miR-222 mimics negative control and high glucose ＋ miR-222 mimics groups. Expression of PARP-1 protein was measured by western blot. Secretory proteins of tPA and PAI-1 in the cell supernatants was detected by ELISA assay. Results Real-time quantitative PCR demonstrated that the expression level of miR-222 mRNA of HMCs in the high glucose group was 2. 537 times that in the normal glucose group at 24 h. The expression of PARP-1 protein of HMCs in the high glucose group was significantly higher than that of normal glucose group（ P〈0. 05）. In the high glucose＋ miR-222 mimics group,the expression level of PARP-1 protein of HMCs was considerably lower than that in the high glucose group（ P〈0. 05）. No statistical significance was noted between the high glucose ＋ miR-222 mimics negative control and the high glucose groups（ P〉0. 05）. Compared with the normal glucose group,tPA and tPA/PAI-1 ratio were significantly decreased,whereas PAI-1 was evidently increased in the high glucose group（ all P〈0. 05）. Compared with the high glucose group,tPA and tPA/PAI-1 ratio were significantly elevated,whereas PAI-1 was significantly decreased in the high glucose ＋ mimics group（ all P〈0. 05）. No statistical significance was observed in tPA,PAI-1 and tPA/PAI-1 ratio between the high-glucose ＋ miR-222 mimics negative control and the high glucose groups（ all P〉0. 05）. Conclusions High glucose can up-regul

关 键 词：微小核糖核酸-222 多聚二磷酸腺苷核糖聚合酶-1 组织型纤溶酶原激活物/低溶酶原激活物抑制物-1 

分 类 号：R587.2[医药卫生—内分泌] R692[医药卫生—内科学]