万古霉素对大鼠肾小管上皮细胞肾损伤定量评价及其机制研究

Quantitative evaluation study of vancomycin-induced injury of cultured renal epithelial cells in rats and its mechanism

目的定量评价不同浓度万古霉素对大鼠肾小管上皮细胞的肾损伤程度,并探讨其可能机制。方法健康雄性SD大鼠肾小管上皮细胞经分离后原代培养,加入含不同浓度万古霉素(0μg/ml、10μg/ml、20μg/ml、30μg/ml、40μg/ml、50μg/ml、60μg/ml)的培养液,3h后测定各组细胞培养基中乳酸脱氢酶(LDH)释放百分率,同时对20μg/ml万古霉素作用浓度的不同时间组(0h、3h、6h、12h、24h、48h)丙二醛(MDA)的含量变化进行考察。结果细胞培养基中,万古霉素作用后的细胞培养上清液MDA含量随着万古霉素作用浓度的升高而增高。与空白对照组相比,20μg/ml组、30μg/ml组、40μg/ml组、50μg/ml组和60μg/ml组的LDH释放百分率均显著增加,差异有统计学意义(P<0.05),细胞上清液MDA含量随着时间延长而增高,与空白对照组相比,24h组和48h组的MDA含量均显著增加,差异有统计学意义(P<0.05)。结论万古霉素对大鼠肾小管上皮细胞具有肾毒性,且呈剂量依赖性,其机制可能与氧化应激反应有关。

Objective To evaluate vancomycin-induced injury of cultured renal epithelial cells in rats quantitatively and explore its mechanism.Methods Renal epithelial cells of healthy rats were isolated and cultured respectively.The cells were stimulated with seven concentrations of vancomycin(0μg/ml,10μg/ml,20μg/ml,30μg/ml,40μg/ml,50μg/ml and 60μg/ml)and the release percentages of lactate dehydrogenase(LDH)were detected 3h later.The contents of malonaldehyde(MDA)in the supernatant of cell culture stimulated with vancomycin concentration of 20μg/ml were detected for different time(0h,3h,6h,12h,24h and 48h).Results The release percentages of LDH and the contents of MDA in the supernatant of cell culture increased with concentrations of vancomycin and time,respectively.Compared with blank group,the release percentages of LDH increased significantly in 20μg/ml group,30μg/ml group,40μg/ml group,50μg/ml group and 60μg/ml group(P<0.05),and the contents of MDA increased significantly in 24h group and 48h group(P<0.05).Conclusion Vancomycin can induce dose dependent injury of cultured renal epithelial cells in rats,and it may be due to oxidative stress mechanism.