PPARα激动剂非诺贝特在高糖诱导的肾小球系膜细胞增殖中的作用  

The role of peroxisome proliferators-activated receptor-α（PPARα） agonist fenofibrate in proliferation of glomerular mesangial cells induced by high glucose

作　　者：熊艳[1,2] 何泳 曾锐[1] 何金森[1] 李维[1] 杨娟[1] 吕倩[1] 李月强[1] 邵菊芳[1] 姚颖[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肾内科,武汉430030 [2]武汉市普爱医院肾内科 [3]武汉市第五医院肾内科

出　　处：《临床肾脏病杂志》2013年第4期157-160,共4页Journal Of Clinical Nephrology

基　　金：国家自然科学基金面上项目（8117（1686）;教育部重大项目[2011]313号（311028）

摘　　要：目的研究过氧化物酶体增殖物激活受体α（PPARα）激动剂非诺贝特在高糖诱导的肾小球系膜细胞中的作用，探讨其对糖尿病肾脏疾病的可能保护作用。方法将。肾小球系膜细胞分为6组：正常对照组（5mmol／L葡萄糖）、高糖组（40mmol／L葡萄糖）和高糖＋不同浓度PPARα激动剂组（FN，40mmol／L葡萄糖加1、10、50、100μmol／L非诺贝特）。应用逆转录-聚合酶链反应（RT-PCR）和Westernblot检测各组肾小球系膜细胞PPARα的表达；通过转染含PPAR反应元件的p4XPPRE-luc重组质粒，双荧光素酶分析法检测体外不同组细胞PPARα活性的改变；MTT法检测细胞增殖；Westernblot检测α平滑肌肌动蛋白（α-SMA）和Ⅳ型胶原蛋白的表达情况。结果高糖刺激组系膜细胞PPARα mRNA及蛋白表达均上调；非诺贝特组上述表达水平则显著升高，且呈浓度依耐性；正常。肾小球系膜细胞弱表达PPARα受体，高糖组PPARα活性无显著升高，而合适浓度的非诺贝特干预后PPARα受体活性明显增加；高糖组Q-SMA及Ⅳ型胶原蛋白表达明显上调，非诺贝特干预后上述指标明显下调。Objective To investigate the role of peroxisome proliferators-activated receptor-α （PPARα） agonist fenofibrate in the proliferation of glomerular mesangial cells （GMCs） induced by high glucose and to study its protective role in the diabetic nephropathy. Methods RT-PCR and West- ern blotting were used to detect the expression of PPARα mRNA and protein in different groups. After transient transfection of an expression vector containing four copies of a consensus PPAR response ele- ment（PPRE） placed upstream from the PGL2-basic luciferase reporter （p4xPPRE-Luc）,dual Lucifer- ase Reporter Gene assay was used to measure the activity of PPARα. MTT assay was used to deter- mine the effect of fenofibrate on cell proliferation. Western blotting was used to examine the expression of α-SMA and Collagen-IV proteins in different groups. Results （1） The mRNA and protein expres- sion of PPARα, and the activity of PPARα were increased in high glucose group and fenofibrate group （dose-dependent） ; （2） The proliferation of GMCs at different time points after incubation with differ- ent concentrations of fenofibrate was statistically different from that of controls; （3）Fenofibrate could dramatically suppress the high-glucose-mediated expression of α-SMA and Collagen-Ⅳ protein. Conclu- sions Fenofibrate inhibited proliferation and the expression of fibronectin,α-SMA and Collagen-Ⅳ of GMCs, which may play a renoprotective role in diabetic nephropathy.

关 键 词：非诺贝特 细胞增殖 过氧化物酶体增殖物活化受体 

分 类 号：R692.6[医药卫生—泌尿科学]