万古霉素联合多黏菌素B用药对肾小管上皮细胞的作用

Effect of vancomycin combined with polymyxin B on the viability of renal tubular epithelial cells

目的研究万古霉素和多黏菌素B联合用药对肾小管上皮细胞(HK2)活力的影响。方法搭建并验证多黏菌素B的人体生理药代动力学(PBPK)模型。基于模型预测万古霉素、多黏菌素B在人体肾脏、肾小管中的组织分布特征。四唑氮氢氧化物法(XTT)检测万古霉素低剂量组(0.5 mg/mL)、万古霉素高剂量组(2.0mg/mL)、多黏菌素组(50 mg/mL)、万古霉素低剂量+多黏菌素组、万古霉素高剂量+多黏菌素组分别对HK2细胞活力及细胞线粒体超氧化物生成的影响。结果建立并验证多黏菌素B PBPK模型,建立及验证的模型所预测的PK参数值与实测PK参数比值均在0.81.2之间。预测多黏菌素B静脉滴注给药后肾脏和肾小管浓度均高于血浆浓度,肾小管药物浓度约为血浆的10倍。肾脏、肾小管药物达峰时间明显滞后,其中肾小管浓度达峰时间为5.3 h,浓度高于50μg/mL的持续时间为8.5 h。细胞活力实验结果显示,8和24 h后万古霉素低剂量组、万古霉素高剂量组、多黏菌素B组、万古霉素低剂量+多黏菌素B组、万古霉素高剂量+多黏菌素B组细胞增殖抑制率分别为1.73%、4.77%、10.34%、10.54%和11.76%及1.95%、7.46%、11.27%、11.55%和16.49%。其中,只有万古霉素高剂量+多黏菌素B组的HK2细胞8 h后产生有统计学意义的线粒体超氧化物。结论万古霉素与多黏菌素B的联合使用可增加肾小管毒性,特别是高剂量万古霉素合用多黏菌素B促使细胞产生线粒体超氧化物。

Objective To investigate the effect of vancomycin combined with polymyxin B on the viability of human renal tubular epithelial cells(HK2).Methods We developed and validated a physiologically based pharmacokinetic model(PBPK)of polymyxin B(PMB).Tissue distribution of vancomycin(VAN)and PMB in the renal tubule and kidney was predicted based on the PBPK model.The effects of VAN and/or PMB[low dose VAN group(0.5 mg/mL),high dose VAN group(2 mg/mL),PMB group(50 μg/mL),low dose VAN + PMB group,high dose VAN + PMB group]on the vitality of HK2 cells and mitochondrial superoxide production were detected by XTT assay.The production of mitochondrial superoxide in HK2 cells was detected by MitoSox.Results The PBPK model of PMB in human was established and validated.The ratio of predicted PK parameters and the observed PK parameters was in the range of 0.8~1.2.It was predicted that the concentration of PMB in the kidney and renal tubules was higher than that in plasma,and the concentration in renal tubules was about 10 times higher than that in plasma.Tmaxof PMB in kidney and renal tubules were significantly delayed.The Tmaxin renal tubules was about 5.3 h,and the duration of concentration higher than 50 μg/mL in renal tubules was 8.5 h.The results of cell viability assay showed that the inhibition rates of cell proliferation in low dose VAN group,high dose VAN group,PMB group,low dose VAN + PMB group,high dose VAN + PMB group were 1.73%,4.77%,10.34%,10.54%,11.76% after 8 h,and 1.95%,7.46%,11.27%,11.55%,and 16.49% after 24 h,respectively.Among them,only the high dose VAN + PMB group produced statistically significant mitochondrial superoxide in HK2 cells.Conclusion The combination of VAN and PMB can increase the toxicity of renal tubules.In particular,high-dose VAN(2.0 mg/mL)+ PMB(50 μg/mL)can promote the production of mitochondrial superoxide in cells.