摘要
目的研究铅对人肾小管上皮细胞(HK-2细胞)的损伤作用,并从氧化应激及线粒体损伤方面探讨其可能的作用机制。方法以不同浓度的乙酸铅处理培养的HK-2细胞,AUTOLAB-18全自动生化分析仪测定细胞上清液乳酸脱氢酶(LDH),硫代巴比妥酸荧光法测定丙二醛(MDA),Annexin V-FITC/PI双染流式细胞仪技术检测细胞凋亡率及坏死率,荧光染料2′,7′-二氯荧光素乙二酯(2′,7-′diehlorofluoreseein diacetate,DCFH-DA)、Monochlorobimane及10-壬基吖啶橙(10-nonyl acridine orange,NAO)结合荧光化学发光仪分别检测细胞内活性氧(ROS)、谷胱甘肽(GSH)及线粒体心磷脂(CL)水平。结果乙酸铅能造成HK-2细胞LDH外漏增加,细胞上清液MDA水平上升,细胞内ROS生成增加,GSH水平改变,线粒体CL氧化损伤,诱发HK-2细胞凋亡及坏死,且凋亡率远较坏死率高。结论氧化应激介导HK-2细胞线粒体损伤,从而启动线粒体途径的细胞凋亡可能是铅致肾细胞毒作用机制之一。
Objective To investigate the possible mechanism about the damage effect in HK-2 cells induced by lead acetate from oxidatation stress and mitochondrial pathway. Methods HK-2 ceils were exposed to different levels of lead acetate in vitro. Actate dehydrogenase and malonaldehyde in cell culture fluid were measured by AUTOLAB-18 automatic biochemistry analyzer and TBA fluorometric method respectively. AnnexinV-FITC/PI double labeled assay was performed by using the flow cytometry methods to detect apoptosis and necrosis. Intracellular reactive oxygen species, glutathione and mitochondrial cardiolipin were detected by Fluoroskan Ascent FL respectively with 2＇, 7＇-diehlorofluoreseein diacetate, monochlorobimane and 10-nonyl acridine orange. Results After exposed to different levels of lead acetate, actate dehydrogenase outleakage from HK-2 cells and malonaldehyde concentration were increased. Generation of ractive oxygen species was enhanced, glutathione level was aherred, mitochondrial cardiolipin was damaged. Cell apoptosis and necrosis were enhanced. Conclusion Mitochondrial apoptosis pathway, which switched on Oxidation stress-mediated mitochondrial damage, may be one possible damage mechanism of HK-2 cells exposed to lead acetate.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2008年第5期359-362,共4页
Journal of Toxicology
关键词
铅
活性氧簇
细胞凋亡
线粒体
Lead
Reactive Oxygen Species
Cell Apoptosis
Mitochondria
