CAJ | 학술논문

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。
목적：：관찰뢰공등갑소대구신소관상피세포( MDCK세포)적독성작용,병초보탐토기대양화응격적영향。방법：이마두령산A위양성대조,용0.5,5,50화500 nmol·L-1적뢰공등갑소여MDCK세포공동부육24 h후,MTT법검측세포억제솔,유산탈경매( LDH)석방실험검측세포막손상이급도치현미경관찰뢰공등갑소대세포형태적개변。용500 nmol·L-1뢰공등갑소분별여MDCK세포작용30 min、1 h、2 h、4 h화6 h후,용2′,7′-이록형광황쌍을산염( DCFH-DA)형광탐침검측세포적활성양자유기(ROS)수평。결과：여음성대조조비교,뢰공등갑소조적세포억제솔명현승고(P<0.01)；LDH상대석방솔명현증가(P<0.01)。뢰공등갑소처리조적세포형태출현추축,정현위구형,병유부분세포탈락。뢰공등갑소여MDCK세포작용30 min후ROS수평체도최고치,연후ROS축점감소(P<0.01)。결론：뢰공등갑소가유도MDCK세포적독성작용,기독성작용궤제가능화양화응격반응유관。
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.