HO-1在造影剂所致肾小管上皮细胞损伤中的作用  

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

作　　者：刘怡晟[1] 朱淳[1] 蒋更如[1] 孙静姝[1] 刘秀英[1] 

机构地区：[1]上海交通大学医学院附属新华医院肾脏内科,200092

出　　处：《放射免疫学杂志》2011年第5期540-544,共5页Journal of Radioimmanology

摘　　要：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 thiazolyltetrazolium,MTT)法测定细胞增殖活力,DCFH-DA荧光探针测定细胞内活性氧(ROS),硫代巴比妥酸法测定细胞内丙二醛(MDA)含量,黄嘌呤氧化酶法测定超氧化物岐化酶(SOD)活性,RT-PCR测定细胞HO-1mRNA表达,Western blot检测细胞HO-1蛋白表达。结果:造影剂碘海醇100mgI/ml可明显抑制HK-2细胞增殖、引起培养液中LDH水平上升、促使细胞内ROS含量增高、MDA含量上升而SOD活性下降,同时诱导HO-1mRNA和蛋白表达上调。与造影剂损伤组相比,HO-1诱导剂CoPP处理组,在引起HO-1表达升高的同时,HK-2细胞增殖增高、培养液LDH水平下降、细胞内ROS含量降低、MDA含量下降而SOD活性上升。HO-1抑制剂ZnPP处理组,在引起HO-1表达下降的同时,HK-2细胞增殖下降、培养液LDH水平升高、细胞内ROS含量减少、MDA含量升高而SOD活性下降。结论:HO-1对造影剂所致肾小管上皮细胞损伤具有明显的保护作用,其作用机制与抑制细胞内氧化应激反应有关。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（P0.01）.The expressions of HO-1mRNA and protein increased in contrast injury group compared with those in normal control group（P0.01）.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（P0.05,P0.01）.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（P0.05,P0.01）.Conclusion HO-1 can protect the tubular epith

关 键 词：血红素加氧酶-1 造影剂 肾小管上皮细胞 活性氧 

分 类 号：R692[医药卫生—泌尿科学]