全反式维A酸对缺氧性肾小管上皮细胞Prohibitin1和Prohibitin2表达的影响  

Effect of all-trans retinoic acid on expressions of Prohibitinl and Prohibitin2 in hypoxic damage of renal tubu- lar epithelial cells

作　　者：徐会凌[1] 覃远汉[1] 周添标[1] 李正一[1] 雷凤英[1] 黄韦芳[1] 

机构地区：[1]广西医科大学第一附属医院儿科,南宁530021

出　　处：《中华实用儿科临床杂志》2013年第5期333-336,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81060061）;广西自然科学基金（0832121）

摘　　要：目的探讨全反式维A酸（ATRA）对缺氧性。肾小管上皮细胞（RTEC）Prohibitinl（PHBl）和Pro—hibitin2（PHB2）表达的影响。方法购自上海生物细胞库的大鼠近端肾小管上皮细胞株NRK-52E，采用混有胎牛血清和双抗的培养基（100mL培养基中加5mL胎牛血清和1mL双抗）在37℃、50mL／L二氧化碳的培养箱中培养。传代3次后分为正常组、模型组和ATRA干预组。正常组细胞不做任何处理，模型组细胞置于真空罐中，负压吸引器吸尽残余空气，充入缺氧气体（950n，L／L氮气和50mL／L二氧化碳）密封建立缺氧性RTEC损伤模型，ATRA干预组细胞中加入0．1μmol／LATRA后参照模型组的方法进行缺氧。于缺氧24h、36h采用实时荧光定量PCR法测定各组NRK-52E细胞PHB1、PHB2、转化生长因子-β1（TGF-β1）的mRNA表达，Westernblot法检测各组NRK-2E细胞PHB1、PHB2、TGF-β1的蛋白表达。结果1．与正常组比较，模型组和ATRA干预组2个时间点NRK-2E细胞的PHB1和PHB2的蛋白表达量及其mRNA表达量均显著下降（P均〈0．05），缺氧时间越长，表达量越低；与模型组比较，ATRA干预组2个时间点NRK一52E细胞的PHB1和PHB2的蛋白表达量及其mRNA表达量均显著上升（P均〈0．05）。2．与正常组比较，模型组和ATRA干预组2个时间点NRK-52E细胞的TGF-β1的蛋白表达量及其mRNA表达量均显著上升（P均〈0．05），缺氧时间越长，表达量越高；与模型组比较，ATRA干预组2个时间点NRK-52E细胞的TGF-β1的蛋白表达量及其mRNA表达量均显著下降（P均〈0．05）。结论ATRA可显著增强缺氧性RTEC中PHB1和PHB2蛋白表达量及其mRNA表达量，可能对缺氧性RTEC有保护作用。Objective To investigate the effect of all-trans retinoie acid（ATRA） on the expressions of Prohi- bitinl （ PHB1 ） and Prohi-bitin2 （ PHB2 ） in hypoxic damage of renal tubular epithelial cells （ RTEC ）. Methods Rat proximal tubular epithelial cell line NRK-52E culture was performed in the 37 ~C 50 mlML carbon dioxide incubator with mixture medium of fetal bovine serum and double-antibody（ 100 mL medium plus 5 mL fetal calf serum and I mL double-antibody）. After 3 times cell propagation, the cells were divided into 3 groups：normal group, model group and ATRA intervention group. The normal group wasn＇t disposed, and the model group was put into vacuum tank filled with hypoxic gas （950 mL/L nitrogen and 50 mL/L carbon dioxide） to induce a hypoxic damage of RTEC. The ATRA inter- vention group was added 0.1 p^mol/L ATRA and was treated with hypoxic gas as model group. After 24 h and 36 h ,the mRNA expressions of PHBI, PHB2 and transforming growth factor-J31 （ TGF-[31 ） were measured by using real-time PCR method,and the protein expressions of PHB1 ,PHB2 and TGF-[31 were detected by using Western blot method. Re- suits 1. Compared with normal group, NRK-52E cells PHB1, PHB2 protein expressions and mRNA expressions at 2 time points （24 h, 36 h） were significantly decreased in model group and ATRA intervention group （all P 〈 0.05 ）, and the longer hypoxia time ,the lower expression value. Compared with model group ,NRK-52E cells PHB1 and PHB2 pro- tein expressions and mRNA expressions in ATRA intervention group were increased significantly at 2 time points （all P 〈 0.05 ）. 2. Compared with normal group, NRK-52E cells TGF-[31 expressions and mRNA expressions at 2 time points（24 h,36 h） were significantly increased in model group and ATRA intervention group（ all P 〈0.05）,and the longer hypoxia time, the higher expression value;Compared with model group, NRK-52E cells TGF-[31 protein expres- sions and mRNA expressions in ATRA intervention group were decreased signific

关 键 词：Prohibitin1 Prohibitin2 全反式维A酸 

分 类 号：R692[医药卫生—泌尿科学]