万古霉素诱导人肾小管上皮细胞损伤及其Smad4表达的研究  

Injury human renal tubular epithelial cell induced by vancomycin and its expression of Smad4

作　　者：高瑞 肖晓琳 许建春[1] 宋玮[1] GAO Rui;XIAO Xiaolin;XU Jianchun;SONG Wei(Department of Pharmacy,The Fifth Central Hospital of Tianjin,Tianjin 300450,China;Central Laboratory,The Fifth Central Hospital of Tianjin,Tianjin 300450,China)

机构地区：[1]天津市第五中心医院药剂科,天津300450 [2]天津市第五中心医院中心实验室,天津300450

出　　处：《现代药物与临床》2024年第1期8-13,共6页Drugs & Clinic

基　　金：天津市滨海新区卫健委科技项目(2019BWKQ029)。

摘　　要：目的 研究万古霉素诱导人肾小管上皮细胞(HK-2)损伤及其Smad同源物4(Smad4)表达,并探讨其潜在的作用机制。方法 使用不同终浓度(0、1、2、3、4、5 mmol/L)的万古霉素溶液作用于HK-2细胞24 h,显微镜下观察万古霉素对细胞形态的影响;蛋白质免疫印迹法检测Smad4、B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)、Bcl-2的蛋白水平;TUNEL法检测HK-2细胞凋亡情况;CCK8法检测HK-2细胞活力来研究万古霉素对肾脏的毒性作用。构建Smad4过表达质粒,通过CCK8法和TUNEL实验检测Smad4过表达对万古霉素诱导的HK-2细胞损伤的影响。结果 与对照组比较,随着万古霉素浓度升高,HK-2细胞逐渐变形,数量逐渐减少;细胞活力显著降低,凋亡率显著增加(P<0.05、0.01)。与对照组比较,促凋亡蛋白Bax表达显著增高,而抗凋亡蛋白Smad4、Bcl-2表达则显著降低(P<0.05、0.01、0.001)。过表达Smad4后,使万古霉素诱导的HK-2的细胞活力显著增加,凋亡率显著降低(P<0.05、0.001)。结论 万古霉素能够诱导人肾小管上皮细胞损伤,其作用机制可能与调控Smad4的蛋白降解密切相关。Objective To study the injury of human renal tubular epithelial cells(HK-2)induced by vancomycin and the expression of Smad4,and to explore the potential mechanism of action.Methods HK-2 cells were treated with vancomycin solution at different final concentrations(0,1,2,3,4,and 5 mmol/L)for 24 h,and the effect of vancomycin on cell morphology was observed under microscope.The protein levels of Smad4,Bax,and Bcl-2 were detected by Western blotting.The apoptosis of HK-2 cells was detected by TUNEL method.The activity of HK-2 cells was detected by CCK8 method to study the toxic effect of vancomycin on kidney.Smad4 overexpression plasmid was constructed,and the effect of Smad4 overexpression on vancomycin-induced HK-2 cell damage was detected by CCK8 assay and TUNEL assay.Results Compared with the control group,with the increase of vancomycin concentration,the number of HK-2 cells was gradually deformed and decreased.The cell viability was significantly decreased and the apoptosis rate was significantly increased(P<0.05,0.01).Compared with the control group,the expressions of pro-apoptotic protein Bax were significantly increased,while the expressions of anti-apoptotic proteins Smad4 and Bcl-2 were significantly decreased(P<0.05,0.01,0.001).After overexpression of Smad4,vancomycin-induced HK-2 cell viability was significantly increased,and apoptosis rate was significantly decreased(P<0.05,0.001).Conclusion Vancomycin can induce the injury of human renal tubular epithelial cells,and its mechanism may be closely related to the regulation of Smad4 protein degradation.

关 键 词：万古霉素 人肾近曲小管上皮细胞 肾损伤 Smad同源物4 B淋巴细胞瘤-2 B淋巴细胞瘤-2相关X蛋白 

分 类 号：R965[医药卫生—药理学]