万古霉素对人肾小管上皮细胞中肾损伤分子-1表达的影响研究

目的:探讨万古霉素对人肾小管上皮细胞(HK-2细胞)中肾损伤分子-1(KIM-1)表达的影响及其可能机制。方法:体外培养HK-2细胞,按万古霉素作用浓度和时间分为不同浓度(0、10、20、30、40、50、60μg·mL-1)和不同时间(0、3、6、12、24、48h)组。显微镜观察HK-2细胞形态学变化,反转录-聚合酶链反应检测KIM-1mRNA表达,细胞免疫荧光法和免疫印迹法检测KIM-1蛋白表达,硫代巴比妥酸法检测HK-2细胞上清液中丙二醇(MDA)的含量。结果:镜下显示,0μg·mL-1组贴壁细胞生长良好,20、40μg·mL-1组部分贴壁细胞脱落和坏死,60μg·mL-1组大部分细胞脱落和坏死;与0μg·mL-1组比较,各浓度组KIM-1mRNA的表达均明显增强(P<0.05或P<0.01),10、20μg·mL-1组KIM-1蛋白表达明显增强(P<0.01),30、40、50、60μg·mL-1组MDA含量明显增强(P<0.01);与0h组比较,24、48h组KIM-1mRNA和蛋白表达、MDA含量均显著增强(P<0.05或P<0.01)。结论:万古霉素刺激可促进HK-2细胞KIM-1mRNA和蛋白的表达,其机制可能与万古霉素致HK-2细胞氧化应激有关。

Effect of Vancomycin on Kidney Injury Molecule-1 Expression in Human Kidney Tubular Epithelial Cell

OBJECTIVE： To explore the effect of vancomycin on kidney injury molecule-1 （KIM-1） expression in human kidney tubular epithelial cell. METHODS： HK-2 cells were cultured in vitro and stimulated with seven concentrations of vancomycin （0, 10, 20. 30, 40, 50, 60 μg·mL-1） for different time （0, 3, 6, 12, 24, 48 h）. The morphology of HK-2 cells was observed under inverted microscope. The mRNA and protein expressions of KIM-1 were detected by RT-PCR and irnmunofluorescence and Western blot, respectively. The contents of MDA in supematant fluid were detected by TBA. RESULTS： Adherent cells grew well in 0μg·mL-1 group, some adherent cells fell off and got necrosis in 20, 40μg·mL-1 group, and most of adherent cells fell off and got necrosis in 60 μg·mL-1 group. Compared with 0 μg·mL-1 group, the mRNA expression of KIM-1 in HK-2 cells was significantly increased （P＇〈0.05 or P〈0.01 ） when treated with different concentrations of vancomycin; the protein expression of KIM-1 in 10, 20 μg·mL-1 groups were increased significantly （P〈0.01）; the Content of MDA increased significantly in 30, 40, 50, 60μg·mL-1 （P〈0.0i）. Compared with 0 h group, the mRNA and protein expression of K1M-1 and MDA content were increased significantly （P〈0.05 or P〈0.01 ）. CONCLUSION： Vancomycin stimulates the mRNA and protein expression of KIM-1 in HK-2 cells, and it may be due to oxidative stress mechanism.