摘要
目的:采用庆大霉素、顺铂、马兜铃酸Ⅰ诱导兔原代肾小管上皮细胞构建体外肾毒性细胞模型。方法:分离培养兔原代肾小管上皮细胞(RTECs),酶化学染色和免疫组化鉴定;采用MTT法绘制兔RTECs传代培养生长曲线并确定最佳接种细胞量及加药时间;测定不同浓度庆大霉素、顺铂、马兜铃酸Ⅰ对细胞增殖作用及乳酸脱氢酶(LDH)释放量的影响。结果:碱性磷酸酶染色及免疫组化染色均呈阳性结果证明获得兔RTECs,细胞传代培养的最佳接种量为1×104个/孔,最佳给药时间为接种48h。不同浓度庆大霉素、顺铂、马兜铃酸Ⅰ均使细胞形态有一定的改变,对细胞增殖均有抑制作用,且呈浓度依赖性。三者均使细胞LDH的释放量增高,呈浓度依赖性,其中顺铂和马兜铃酸Ⅰ的作用最显著。结论:采用庆大霉素、顺铂、马兜铃酸Ⅰ可诱导兔原代肾小管上皮细胞构建体外肾毒性细胞模型。
Objective： To build the cellular model of renal toxicity in vitro by gentamicin,cisplatin and aristolochic acid Ⅰinduced the rabbit primary renal tubular epithelial cells. Methods： Rabbit primary renal tubular epithelial cells（ RTECs） were isolated and identified by enzymechemical staining and immunohistochemistry. The growth curve of rabbit RTECs subculture was drawn and the optimum inoculation cell quantity and administration time were confirmed by MTT method. The effects,including the cell proliferation and lactate dehydrogenase（ LDH） release by gentamicin,cisplatin,aristolochic acid Ⅰof different concentration,were determined. Results： The staining of alkaline phosphatase and immunohistochemical result showed positive results,which proved that RTECs were cultured successfully. The optimum inoculation cell quantity was 1 × 104 cells / well and the optimum administration time was 48 h after subculture. Gentamicin,cisplatin and aristolochic acid Ⅰof different concentration could change the cell morphology and inhibit the cell proliferation with a slightly concentration dependence manner.Three kinds of xenobiotics increased the LDH release of RTECs with a concentration dependence manner,the effects of cisplatin and aristolochic acid Ⅰ were significant. Conclusion： Using gentamicin,cisplatin and aristolochic acid Ⅰinduce the rabbit primary renal tubular epithelial cells could build the cellular model of renal toxicity in vitro.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2016年第1期209-213,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
贵州大学引进人才科研项目(贵大人基合字(2012)010号)
贵州大学研究生创新基金项目(研农2015020)
贵州省国际科技合作计划项目(黔科合外G字[2013]7009号)
贵州省科学技术基金项目(黔科合J字[2012]2173号)
关键词
庆大霉素
顺铂
马兜铃酸Ⅰ
肾小管上皮细胞
肾毒性
gentamicin
cisplatin
aristolochic acid Ⅰ
renal tubular epithelial cell
renal toxicity
作者简介
通讯作者
