【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(5 mmol/L葡萄糖)、高糖组(40 mmol/L葡萄糖)和高糖+不同浓度PPARα激动剂组(FN,40 mmol/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(RTPCR)和Western blot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;Western blot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PPARαmRNA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α-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。结论 非诺贝特可通过抑制高糖诱导的肾小球系膜细胞增殖及细胞外基质蛋白的表达而对糖尿病肾脏疾病起保护作用更多还原

【Abstract】 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 Western 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 element(PPRE) placed upstream from the PGL2-basic luciferase reporter(p4xPPRE-Luc),dual Luciferase Reporter Gene assay was used to measure the activity of PPARα.MTT assay was used to determine 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 expression 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 different 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.Conclusions Fenofibrate inhibited proliferation and the expression of fibronectin,α-SMA and Collagen-Ⅳ of GMCs,which may play a renoprotective role in diabetic nephropathy.更多还原