内质网应激在对比剂诱导肾小管上皮细胞凋亡中的作用  

Effect of Endoplasmic Reticulum Stress on the Apoptosis of Contrast Media-Induced Renal Tubular Cells

作　　者：杨一菲[1] 杨定平[1] 贾汝汉[1] 丁国华[1] 

机构地区：[1]武汉大学人民医院肾内科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2013年第4期507-511,共5页Medical Journal of Wuhan University

基　　金：湖北省自然科学基金资助项目(编号:2007ABA254)

摘　　要：目的:探讨内质网应激(ERS)在对比剂诱导的肾小管上皮细胞凋亡中的作用及N-乙酰半胱氨酸(NAC)的保护作用。方法:将不同浓度(50,100,150mgI/ml)碘普罗胺分别加入体外培养的大鼠肾小管上皮细胞(NRK-52E)中孵育1h;10mmol/L NAC预处理细胞1h后,加入100mgI/ml碘普罗胺共孵育1h。Hoechst-33342染色法检测肾小管上皮细胞凋亡率;荧光探针检测细胞内活性氧(ROS)的产生;Western印迹法检测细胞内内质网功能调节蛋白GRP78、内质网源性转录因子CHOP表达水平。结果:对比剂呈浓度依赖性诱导NRK-52E细胞凋亡,且呈浓度依赖性诱导NRK-52E细胞内ROS生成及GRP78、CHOP蛋白表达上调(P<0.05);经NAC预处理后,细胞内ROS生成及细胞凋亡率明显下降(P<0.05),且GRP78、CHOP蛋白表达下调(P<0.05)。结论:碘普罗胺能诱导肾小管上皮细胞凋亡,其机制可能与细胞内ROS生成增加,诱发ERS,上调GRP78、CHOP蛋白表达有关。抗氧化剂NAC通过降低NRK-52E细胞内ROS介导的ERS,减少肾小管上皮细胞凋亡。Objective： To investigate the effect of endoplasmic reticulum stress （ERS） on contrast media-induced renal tubular cell apoptosis and evaluate the effect of N-acetylcysteine （NAC） on it. Methods. NRK-52E cells were exposed to different concentrations of iopromide （50, 100 and 150 mg/ml） for 1 hour. Cells of other groups were treated with NAC （10 mmol/L） for 1 hour firstly and then incubated with iopromide （100 mgI/ml） for 1 hour; Hoechst 33342-staining was used to measure the apoptotic rate of renal tubular ceils; Fluorescent probe CM-HZDCFDA was used to evaluate the level of reactive oxygen species （ROS） ; Western blot was used to test the ex- pression of GRP78 and CHOP. Results： Iopromide induced NRK-52E cells apoptosis in a concen- tration-dependant manner and increased the levels of ROS, GRP78 and CHOP in a concentration- dependent manner （P〈0.05）. After the pretreatment of NAC, the level of ROS and apoptosis decreased （P〈0.05）, and the expression of GRP78 and CHOP were also reduced significantly （P〈0. 05）. Conclusion.. Iopromide could induce the apoptosis of NRK-52E cells. The possible mechanism may he explained by the increase of intracellular ROS, which may trigger the ERS and upregulate the expression of GRP78 and CHOP proteins. NAC belongs to the group of antioxida- nts, and it could decrease the level of ROS in NRK-52E cells, weaken the ERS and finally reduce the apoptosis.

关 键 词：对比剂 内质网应激 肾小管上皮细胞 细胞凋亡 活性氧 N-乙酰半胱氨酸 

分 类 号：R34[医药卫生—基础医学] R692