Prohibitin1和Prohibitin2在缺氧性肾小管上皮细胞损伤中的表达及作用

Expression and Function of Prohibitin1 and Prohibitin2 in Hypoxic Damage of Renal Tubular Epithelial Cells

目的探讨Prohibitn1(PHB1)、Prohibitin2(PHB2)在缺氧性肾小管上皮细胞(RTEC)损伤中的表达和作用。方法以体外培养的大鼠近端肾小管上皮细胞株(NRK-52E)为对象,NRK-52E细胞置于50 mL.L-1二氧化碳(CO2),37℃培养箱中孵育至80%,传代后随机分为正常组和模型组。正常组细胞继续培养,模型组细胞置于真空罐中,负压吸引器抽尽残余空气,充以配好的缺氧气体(950 mL.L-1氮气和50 mL.L-1CO2)密封建立缺氧性RTEC损伤模型,分别于造模第12、24、36小时采用实时荧光定量PCR检测PHB1、PHB2、转化生长因子-β1(TGF-β1)mRNA表达,Western blot检测PHB1、PHB2蛋白表达。结果 1.与正常组比较,模型组各时间点NRK-52E细胞的PHB1、PHB2蛋白及其mRNA表达均降低(Pa<0.05),缺氧时间越长,表达量越低;NRK-52E细胞的TGF-β1 mRNA表达均增高(Pa<0.05),缺氧时间越长,表达量越高。2.相关性分析:模型组NRK-52E细胞的PHB1、PHB2 mRNA表达与TGF-β1 mRNA表达均呈负相关(r=-0.97、-0.99,Pa<0.05)。结论缺氧所致NRK-52E细胞的PHB1、PHB2蛋白及mRNA表达均降低,缺氧时间越长,表达量越低,NRK-52E细胞损伤越重。PHB1、PHB2可能参与RTEC损伤的发生发展。

Objective To explore the expression and function of Prohibitin1（PHB1） and Prohibitin2（PHB2） in hypoxic damage of renal tubular epithelial cells（RTEC）. Methods Rat′s proximal tubular epithelial cell line（NRK-52E）was performed in vitro for this study.NRK-52E was subcultured and randomly assigned into normal group and model group,until it was growth to 80%.The normal group was placed in incubator under the condition of 50 mL·L-1 carbon dioxide,37 ℃.The model group was established in vacuum tank,the residual air in tank was extracted by suction device and the vacuum tank was filled the gas which composed 950 mL·L-1 nitrogen and 50 mL·L-1 carbon dioxide.After the treatment,the relevant indexes were detected at the 12th,24th and 36th hours,respectively.The expressions of mRNA of PHB1,PHB2,transforming growth factor-β1（TGF-β1） in NRK-52E cells were detected by real-time fluorescent quantitative polymerase chain reaction（PCR）,and Western blot was performed to detect the protein expression of PHB1 and PHB2. Results 1.Compared to normal group,at each same time point,the expressions of protein and mRNA of PHB1 and PHB2 in NRK-52E cells in model group were all decreased（Pa0.05）,the longer the hypoxia,the lower the expression level;The expressions of mRNA of TGF-β1 in NRK-52E cells in model group were all increased（Pa0.05）,the longer the hypoxia,the higher the expression level.2.Correlation analysis showed that：The expressions of mRNA of PHB1 and PHB2 in NRK-52E cells were both negatively correlated with the expression of TGF-β1 mRNA（r=-0.97,-0.99,Pa0.05）. Conclusions The expressions of PHB1 and PHB2 are all decreased in model group,the longer the hypoxia,the lower the expression level,the more severe the degree of damage of NRK-52E cells.PHB1 and PHB2 may participate in the development of hypoxic damage of RTEC.