HO-1在造影剂所致肾小管上皮细胞损伤中的作用

目的:探讨血红素加氧酶-1(Heme oxygenase-1,HO-1)在造影剂所致肾小管上皮细胞损伤中的作用.方法:体外培养的HK-2细胞,分为正常对照组、甘露醇对照组、造影剂损伤组、钴原卟啉(CoPP)处理组、锌原卟啉(ZnPP)处理组.比色法测定细胞培养上清液乳酸脱氢酶(LDH)水平,四甲基偶氮唑(methyl ...

HO-1 Expression on the Renal Tubular Epithelial Cell Injury Induced by Contrast Media

Objective To investigate the expression of HO-1 on the renal tubular epithelial cell injury induced by contrast media.Methods Cultured human proximal tubular epithelial cells(HK-2) were divided into 5 groups: normal control group in which cells treated with serum-free DMEM-F12 medium;mannitol osmotic pressure control group treated with 1.6% mannitol;contrast medium injury group treated with 100mgI/ml Iohexol,CoPP treatment group treated with Iohexol 100mgI/ml and 10μmol/L CoPP,and ZnPP treatment group treated with Iohexol 100mgI/ml and 20μmol/L ZnPP.LDH level in the cell culture media was tested by spectrophotometric method and cell viability was measured by methyl thiazolyl tetrazolium(MTT) method.DCFH-DA fluorescent probe was used to detect reactive oxygen species(ROS) in cells.MDA and SOD in cells were detected by TBA and xanthine oxidase methods.HO-1mRNA and protein expression were determined by RT-PCR and Western blot respectively.Results LDH level in cell culture media,ROS and MDA of HK-2 cells increased whereas the cell viability and SOD activity decreased significantly in contrast medium injury group compared with those in normal control group(P<0.01).The expressions of HO-1mRNA and protein increased in contrast injury group compared with those in normal control group(P<0.01).There was no significant difference between mannitol control group and normal control group in above parameters.In CoPP treatment group,with the increase of HO-1mRNA and protein expression,LDH level in cell culture media,ROS and MDA of HK-2 cells decreased whereas the cell viability and SOD activity increased significantly compared with those in contrast medium injury group(P<0.05,P<0.01).In ZnPP treatment group,with the decrease of HO-1mRNA and protein expression,LDH level in cell culture media,ROS and MDA of HK-2 cells increased whereas the cell viability and SOD activity decreased significantly compared with those in contrast medium injury group(P<0.05,P<0.01).Conclusion HO-1 can protect the tubular epithelial cell injury induced by contrast media,the protective effects related to its anti-oxidant effect.