高糖对体外培养的足细胞自噬的影响  

Effects of high glucose on autophagy of cultured podocytes in vitro

作　　者：郑海宁[1] 苏东明[2] 董成龙[1] 鲁一兵[1] 丁大法[1] 

机构地区：[1]南京医科大学第二附属医院内分泌科,江苏南京210011 [2]南京医科大学代谢病研究中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2014年第4期433-436,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金面上项目(81270896);国家自然科学基金青年项目(81100577)

摘　　要：目的:研究不同葡萄糖浓度、不同作用时间对体外培养的足细胞自噬的影响。方法:体外培养小鼠永生性足细胞系,给予不同浓度的葡萄糖(5.6、11.0、20.0、30.0 mmol/L)刺激,作用时间分别为6、12、24、36 h。Western blot检测足细胞自噬标记蛋白微管相关蛋白1轻链3(LC3)Ⅱ的表达;透射电镜观察足细胞自噬小体的形成;荧光显微镜观察足细胞转染GFP-LC3荧光蛋白后绿色荧光蛋白颗粒的变化。结果:①与0 h相比,30 mmol/L葡萄糖刺激后足细胞LC3-Ⅱ蛋白的表达随着作用时间的延长逐渐增加,到24 h时增加最明显,差异有显著性(P<0.05)。24 h时,透射电镜下足细胞自噬小体形成最多;荧光显微镜下足细胞转染GFP-LC3绿色荧光蛋白颗粒最多。②与对照组5.6 mmol/L相比,随着葡萄糖刺激浓度的增加足细胞LC3-Ⅱ蛋白的表达明显增加,30 mmol/L浓度时最明显,差异有显著性(P<0.05)。结论:高糖呈浓度和时间依赖性增加体外培养的足细胞LC3-Ⅱ蛋白的表达、诱导足细胞自噬体的形成。Objective：To explore the effect of different concentrations of D-glucose and different periods of time on autophagy in in vitro cultured podocytes. Methods：In vitro,cultured mouse podocyte clones （MPC5） were incubated with D-glucose at concentrations of 5.6, 11,20 and 30 mmol/L for 6, 12,24 and 36 h,respeetively. The protein expressions of LC3 in podoeytes were examined by Western blot and autophagosomes in podoeytes were observed by transmission electron microscope. The changes of green fluorescent protein particles were observed after GFP-LC3 transfected podocytes by fluorescence microscopy. Results： （1）Compared with the 0 h group, the protein expression of LC3-II increased in a time-dependent manner and reach the highest at 24 h in podocytes exposed to 30 mmol/L D-glucose （P 〈 0.05）. At the time of 24 h,the autophagosomes in podocytes and the green fluorescent protein particles in GFP-LC3 transfected podocytes were increased most prominent. （2） Compared with the 5.6 mmol/L group,the protein expression of LC3-II was increased in a dose-dependent manner in podocytes exposed to D-glucose with increasing concentrations. In the concentration of 30 mmol/L,the expression of LC3-II was most prominent （P 〈 0.05）. Conclusion：High glucose upregulates the protein expression of LC3-II in time- and dose-dependent manners and induces autophagy in cultured podocytes.

关 键 词：足细胞 自噬 葡萄糖 糖尿病肾病 

分 类 号：R587.2[医药卫生—内分泌]