丹参注射液对造影剂诱导的肾小管上皮细胞损伤的保护作用

目的：研究丹参注射液对造影剂碘海醇诱导的肾小管上皮细胞损伤的保护作用并对其作用机制进行探讨。方法：体外培养的人肾小管上皮细胞（HK-2细胞）,分为正常对照组、甘露醇对照组（与碘海醇渗透压相当）、造影剂对照组（碘海醇100mg/ml）、丹参防治组（碘海醇100mg/ml＋丹参注射液15mg/ml）。比色法测定细胞培养上清液LDH水平,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MDA含量,黄嘌呤氧化酶法测定SOD活性,RT-PCR测定细胞内血红素加氧酶-1（HO-1）mRNA表达,Westernblot检测细胞内HO-1蛋白表达。结果：碘海醇100mg/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和MDA含量增高而细胞内SOD活性下降,同时碘海醇可诱导HO-1mRNA和蛋白表达上调。加入丹参可明显降低培养液中LDH水平,促进细胞增殖,同时抑制细胞内ROS的产生,降低细胞内MDA含量,保护细胞内SOD活性,并使HO-1mRNA和蛋白表达进一步增强。结论：丹参对造影剂碘海醇所致肾小管上皮细胞具有明显的保护作用,其作用机制与其抗氧化能力及诱导HO-1表达有关。

Protective Effects of Salvia Miltiorrhiza Injection on the Renal Tubular Epithelial Cell Injury Induced by Contrast Media

Objective：To investigate the protective effects of Salvia Miltiorrhiza injection on the renal tubular epithelial cell injury induced by contrast media.Methods：Cultured human proximal tubular epithelial cells（HK-2）were divided into 4 groups：normal control group in which cells treated with serum-free DMEM-F12 medium;mannitol osmotic pressure control group treated with 8% mannitol;contrast medium control group treated with 100mgI/ml Iohexol,and Salvia Miltiorrhiza protective group treated with Iohexol 100mgI/ml and Salvia Miltiorrhiza injection 15 mg/ml.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 HO-1 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 viability and SOD activity of HK-2 cells decreased significantly in contrast medium control group compared with those in normal control group（P0.01）.The expression of HO-1mRNA and HO-1 protein increased in contrast control 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 Salvia Miltiorrhiza protective group,LDH level in cell culture media,ROS and MDA of HK-2 cells decreased whereas the viability and SOD activity of HK-2 cells increased significantly compared with those in contrast medium control group（P0.05,P0.01）.The expression of HO-1 mRNA and HO-1 protein increased in Salvia Miltiorrhiza protective group compared with those in contrast control group（P0.05）.Conclusion：Salvia Miltiorrhiza injection can protect the tubular epithelial cell injury induced by contrast media,the protective effects related to its anti-oxidant effect and induction of HO-1.