白藜芦醇对高糖状态下大鼠肾小管上皮细胞增殖、凋亡的影响及机制
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国家自然科学基金资助项目(81270896),南京医科大学重点科研基金项目(2010NJMUZ50)


Effects of resveratrol on proliferation and apoptosis of rat renal tubular epithelial cells under high glucose and underlying mechanisms study
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    目的:观察白藜芦醇对高糖状态下大鼠肾小管上皮细胞NRK-52E增殖-凋亡的影响,并探讨其可能的作用机制。方法:体外培养大鼠NRK-52E细胞,分为正常对照组(NG组,正常葡萄糖5.6 mmol/L)-NG+白藜芦醇(Res)组(NG+Res组,白藜芦醇20 μmol/L)-高糖组(HG组,葡萄糖30 mmol/L)和高糖+白藜芦醇组(HG+Res组,白藜芦醇20 μmol/L),白黎芦醇预处理6 h,加HG刺激后24 h,四甲基偶氮唑盐微量酶反应比色(MTT)法观察肾小管上皮细胞增殖的改变;流式细胞仪检测细胞凋亡;Western blot检测蛋白烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)-葡萄糖调节蛋白78(GRP78)蛋白表达变化。结果:①与NG组相比,高糖能明显增加肾小管上皮细胞的增殖和凋亡;与HG组相比,加用白藜芦醇干预能明显抑制肾小管上皮细胞的增殖和凋亡,差异均具有显著性(P值均 < 0.01);②与NG组相比,高糖刺激NRK-52E细胞24 h后NOX4和GRP78蛋白表达明显增加(P值均 < 0.05);与HG组相比,HG加白藜芦醇干预后NRK-52E细胞NOX4和GRP78蛋白表达量明显减少,差异均具有统计学意义(P值 < 0.05)。结论:白藜芦醇可能通过降低肾小管上皮细胞内氧化应激和内质网应激的产生,减少肾小管上皮细胞的增殖和凋亡而对糖尿病肾病起保护作用。

    Abstract:

    Objective:To investigate the effect of resveratrol on the proliferation and apoptosis of rat renal tubular epithelial cells (NRK-52E) and its possible mechanism. Methods: The NRK-52E cells were cultured in vitro. The NRK-52E cells were divided into four groups: normal glucose (5.6 mmol/L, the NG group), NG plus resveratrol (20 μmol/L, the NG+Res group), high glucose (30 mmol/L, the HG group),and HG plus resveratrol at 20 μmol/L (the HG+Res group). After pretreatment of resveratrol for 6 h and stimulation of HG for 24 h, the rate of cell proliferation was examined by the MTT method, the rate of cell apoptosis was determined by flow cytometry and the expression of GRP78 proteins in the NRK-52E cells was analyzed by Western blot. Results: Compared to the NG group, the rate of cell proliferation and apoptosis in the HG group was increased significantly. Simultaneous incubation with resveratrol inhibited NRK-52E cell proliferation and apoptosis (all P value < 0.01). As compared to the NG group, the expression of NOX4 and GRP78 protein in the HG group was significantly increased (all P value < 0.05). The expression of NOX4 and GRP78 protein in the HG+Res group was significantly decreased than that in the HG group (all P value < 0.05). Conclusion: This data demonstrate that resveratrol may exert antiproliferative and anti-apoptosis effects on rat renal tubular epithelial cells by inhibiting oxidative stress and endoplasmic reticulum stress.

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游 娜,王雅云,董成龙,冯亚敏,缪 珩,丁大法,鲁一兵,刘 超.白藜芦醇对高糖状态下大鼠肾小管上皮细胞增殖、凋亡的影响及机制[J].南京医科大学学报(自然科学版),2015,(4):485-489

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  • 收稿日期:2014-12-11
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  • 在线发布日期: 2015-05-05
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