DJ-1／Nrf2通路对高糖诱导大鼠肾小管上皮细胞损伤的保护作用  

Protective effect of DJ - 1/Nrf2 pathway on rat renal proximal tubular epithelial cell injury induced by high glucose

作　　者：沈孜颖 夏中元[1] 孙倩[1] 赵博[1] 孟庆涛[1] 雷少青[1] 

机构地区：[1]武汉大学人民医院麻醉科,430060

出　　处：《中华实验外科杂志》2015年第12期2993-2995,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81471844）

摘　　要：目的观察高糖诱导大鼠肾小管上皮细胞NRK-52E应激损伤及DJ—1／Nrf2表达的变化，探讨其内源性抗氧化损伤机制。方法大鼠肾小管上皮细胞分别用5．5mmol／L葡萄糖（低糖组）和30mmol／L葡萄糖（高糖组）刺激0、24、48、72、96h。细胞计数试剂盒（CCK-8）实验检测不同时间点下肾小管上皮细胞的损伤程度，丙二醛（MDA）、超氧化物歧化酶（SOD）检测细胞的氧化应激水平，Westernblot检测DJ-1、Nrf2和血红素氧合酶-1（HO-1）蛋白的表达。过表达DJ-1，分为空质粒组和DJ-1过表达组，高糖刺激下观察肾小管上皮细胞凋亡及Nrf2、HO-1蛋白表达水平。结果随着高糖的刺激，48、72、96h时间点的高糖组细胞应激损伤（0．89±0．02、0．82±0．02、0．80±0．03）呈时问依赖性加重，MDA含量[（0．17±0．01）、（0．18±0．03）、（0．23±0．01）nmol／mg蛋白]递进性升高而SOD活性[（29．43±3．35）、（23．95±2．69）、（20．68±1．86）U／mg蛋白]呈时间依赖性下降（P〈0．05）；与0h时间点比较，高糖组48、72h时间点的DJ-1蛋白表达水平（1．63±0．19、1．80±0．16）显著升高，72h时间点的Nrf2和HO-1蛋白表达水平（1．60±0．13、1．83±0．27）亦明显升高（P〈0．05）；然而，与72h时间点比较，96h时间点的DJ-1、Nff2和HO-1蛋白表达水平（1．29±0．10、1．13±0．12、1．22±0．11）显著下降（P〈0．05）。过表达DJ-1可轻度减少高糖引起的细胞损伤（空质粒高糖组：12．19±0．96；DJ-1过表达组：7．31±0．29），并显著增加Nrf2和HO-1蛋白的表达水平（2．20±0．34、1．87±0．31，P〈0．05）。结论DJ-1／Nrf2通路可能参与大鼠肾小管上皮细胞高糖氧化应激中的内源性细胞保护机制。Objective To investigate oxidative stress injury of rat renal proximal tubular epithelial （NRK- 52E） cells under high glucose condition and its antioxidant effect. Methods NRK -52E cells were individually treated with low glucose （ 5.5 mmol/L D - glucose ） and high glucose （ 30 mmol/L D -glucose） for 0, 24, 48, 72 and 96 h. Cell counting kit -8 （CCK -8） test was taken to evaluate cell viability. Malondialdehyde （MDA） contents and superoxide dismutase （SOD） activity were determined to evaluate oxidative stress. Western blotting was adopted to determine the expression of DJ - 1, Nrf2 and heine oxygenase - 1 （ HO - 1 ）. After overexpression of DJ - 1 in NRK - 52E cells, cell apoptosis and ex- pression of Nrf2 and HO - 1 proteins were determined. Results With the stimulation of high glucose, cell injury was time -dependently aggravated in high glucose group at the time points of 48, 72, 96 h （0. 89 ± 0.02, 0.82±0.02, and0.80±0.03）, and the content of MDA [（0.17±0.01）, （0.18±0.03）, and （0. 23± 0. 01 ） nmol/mg pro ] was increased, while the activity of SOD [ （29.43± 3.35 ）, （23.95 ± 2. 69 ）, and （20. 68± 1.86 ） U/rag pro ] was significantly decreased （ P 〈 0. 05 ）. As compared with the 0 - h time point, the expression of DJ - 1 protein in high glucose group was significantly increased at the time points of 48 and 72 h （1.63±0.19, and 1.80±0.16）, and that of Nrf2 and HO-1 proteins （ 1.60 ±0. 13, and 1.83 ± 0. 27） at 72 h was also increased sharply （ P 〈 0. 05）. However, the expres- sion of DJ - 1, Nrf2 and HO - 1 proteins （ 1.29 ±0. 10, 1.13±0. 12, and 1.22 ±0. 11 ） was significantly decreased at 96 h compared to that at 72 h （ P 〈 0. 05 ）. The over - expression of DJ - 1 decreased the cell injury induced by high glucose （7. 31±0. 29 ） and significantly increased the expression of Nrf2 and HO - 1 proteins （2. 20 ±0. 34, and 1.87± 0. 31, P 〈 O. 05 ）. Conclusion The activation of DJ - 1/Nrf2 pathway might par

关 键 词：DJ-1 Nrf2 抗氧化应激 肾近端小管上皮细胞 高糖 

分 类 号：R692[医药卫生—泌尿科学]