马来酸罗格列酮抑制糖基化终产物诱导的人肾系膜细胞RANTES高表达

Rosiglitazone maleate inhibits the hyperexpression of RANTES induced by advanced glycosylation end products in human renal mesangial cells (HRMC)

目的探讨马来酸罗格列酮对糖基化终产物(AGEs)诱导的人肾系膜细胞(HRMC)趋化因子RANTES高表达的影响,及其对糖尿病大鼠血清糖基化终产物-肽(AGE-P)及肾脏RANTES表达的影响。方法运用糖基化修饰的牛血清白蛋白(AGE-BSA)与马来酸罗格列酮干预体外培养的HRMC,ELISA法检测细胞培养上清中RANTES蛋白水平,实时定量RT-PCR检测细胞中RANTES mRNA水平,Western blot检测RANTES蛋白表达。制备2型糖尿病大鼠模型,马来酸罗格列酮治疗10周。流动注射分析法测定血清AGEP,免疫组织化学检测肾皮质RANTES表达。结果马来酸罗格列酮干预组HRMCRANTES的mRNA和蛋白表达以及蛋白分泌明显低于AGE-BSA组;马来酸罗格列酮治疗组糖尿病大鼠血清AGE-P明显下降,肾皮质RANTES表达明显低于糖尿病非治疗组。结论马来酸罗格列酮可以抑制AGE-BSA诱导的HRMCRANTES的表达和分泌,还可降低糖尿病大鼠血清AGE-P及肾脏RANTES表达。

Objective To investigate the effects of rosiglitazone on the upregulation of expression of cbemokine RANTES （regulated upon activation, normal T cell expressed and secreted） induced by advanced glycosylation end products （AGEs） in the cultured human renal mesangial ceils （HRMC）. Methods AGE-BSA was prepared by incubation of bovine serum albumin （BSA） with glucose at 37℃ for 12 weeks. HRMC was cultured in the presence of AGE-BSA （glucose at 50 mmol/L） with rosiglitazone. RANTES mRNA was analyzed by quantitative real time RT-PCR. The contents of RANTES in the cultured supernatant and cell lysates were detected by using ELISA and Western blot, respectively. Diabetic rats induced by streptozotocin were treated with or without rosiglitazone. The AGE-Peptide level in serum was measured by flow injection assay. Immunohistochemistry was applied to detect the expression of RANTES in renal cortex of rats. Results Group of AGE treated with rosiglitazone （RSG） at 0. 1,1, 10,100μmol/L versus AGE without RSG showed the inhibited effects on RANTES mRNA （0. 22,0. 20,0. 14,0.12 vs 1. 45,P〈0.05-0.01）. Serum level of AGE-Peptiede was higher in DM versus control（2.87±0.21 vs 0. 90±0.13U/ml, P〈0.01）, and lower in DM-b RSG than DM （1.45 ± 0.15 vs 2.87 ± 0. 21U/ml, P〈0.01）. The RANTES（＋） particles number each glomerulus was more in DM versus control（8. 97±0.9 vs0. 934-0. 6,P〈0.05） ,and less in DM＋RSG than DM（5. 014±0. 6 vs 8. 974±0.9,P 〈0.05）. Conclusions Rosiglitazone significantly inhibits the expression and secretion of RANTES induced by AGE-BSA in vivo and in vitro.