基于狗肾小管上皮细胞的雷公藤甲素体外肾毒性研究  

Studies on Nephrotoxicity Induced by Triptolide on MDCK Cell Model in vitro

作　　者：阮浩澜 陈琪 黎旸 许姿敏 翁森 

机构地区：[1]广东省食品药品检验所药理毒理室,广州510180

出　　处：《中国药师》2015年第1期1-4,共4页China Pharmacist

基　　金：广东省2011年建设中医药强省科研课题(编号:20111153);广东药检系统科研项目(编号:ZA20101517)

摘　　要：目的:观察雷公藤甲素对狗肾小管上皮细胞(MDCK细胞)的毒性作用,并初步探讨其对氧化应激的影响。方法:以马兜铃酸A为阳性对照,用0.5,5,50和500 nmol·L-1的雷公藤甲素与MDCK细胞共同孵育24 h后,MTT法检测细胞抑制率,乳酸脱氢酶(LDH)释放实验检测细胞膜损伤以及倒置显微镜观察雷公藤甲素对细胞形态的改变。用500 nmol·L-1雷公藤甲素分别与MDCK细胞作用30 min、1 h、2 h、4 h和6 h后,用2',7'-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针检测细胞的活性氧自由基(ROS)水平。结果:与阴性对照组比较,雷公藤甲素组的细胞抑制率明显升高(P<0.01);LDH相对释放率明显增加(P<0.01)。雷公藤甲素处理组的细胞形态出现皱缩,呈现为球形,并有部分细胞脱落。雷公藤甲素与MDCK细胞作用30 min后ROS水平达到最高值,然后ROS逐渐减少(P<0.01)。结论:雷公藤甲素可诱导MDCK细胞的毒性作用,其毒性作用机制可能和氧化应激反应有关。Objective：To study the nephrotoxicity induced by triptolide （ TP） on MDCK cell model and investigate its effect on oxidative stress. Methods：Aristolochic acid was chosen as the positive control. After the MDCK cells were incubated with 0. 5, 5, 50 and 500 nmol.L-1 TP for 24h, MTT method was used to observe the cell inhibiting rate and lactate dehydrogenase （LDH） release test was used to detect the cell membrane damage caused by TP. The cell morphology was observed under an inverted microscope. After the MDCK cells were incubated with 500 nmol.L-1 TP respectively for 30min, 1h, 2h, 4h and 6h, the level of reactive oxygen species （ ROS） was detected using 2′,7′-dichlorodihydro-fluorescein diacetate （ DCFH-DA） as the fluorescent probe. Results：Compared with those of the negative control group, the cell inhibiting rates and the relative LDH release rates in TP-treated group were increased sig-nificantly（P〈0. 01）. The cells in TP-treated group were creased, turned into the round shape and began to shed off. After the MDCK cells were incubated with TP for 30min, the level of ROS reached the highest value, and then began to decrease （P〈0. 01）. Conclu-sion：TP can induce the toxic effects on MDCK cells and the mechanism may be related to oxidative stress.

关 键 词：雷公藤甲素 狗肾小管上皮细胞 毒性 氧化应激 

分 类 号：R285.5[医药卫生—中药学]