碘过量（10-4mol--L）对FRTL细胞早期的影响.docVIP

碘过量（10-4mol/L）对FRTL细胞早期的影响 李敏，李兰英，姚小梅﹡ 摘要：目的：探讨碘过量对Fischer大鼠甲状腺细胞(FRTL) 早期的影响。方法：10-4mol/L 碘化钾（KI）处理FRTL细胞2h、4h、24h，利用MitoSOX通过流式细胞术和荧光显微镜检测线粒体超氧化物生成，免疫细胞化学法检测细胞色素C（cyt c）释放，比色法检测细胞培养液上清乳酸脱氢酶（LDH）活力，利用碘化丙啶通过流式细胞术检测死亡细胞百分比，DNA电泳检测凋亡DNA片段。结果：处理2h、4h、24h，FRTL细胞线粒体超氧化物生成均升高，以2h升高最明显；cyt c从线粒体释放至胞浆，以4h最明显；随处理时间延长，培养液上清LDH活力明显增加（p0.05），死亡细胞百分比升高；处理24h，出现DNA ladder。结论：碘过量（10-4mol/L）处理2h可引起FRTL细胞线粒体过氧化损伤，24h可引起细胞凋亡。 关键词：碘，甲状腺，细胞，培养的，超氧化物类，线粒体 Effects of iodide excess (10-4mol/L) on FRTL cells in early period LI Min，LI Lan-yingYAO Xiao-mei3﹡，1Xushui Peoples Hospital, Hebei Province, China, 072550 ,2Key Lab of Hormones and Development Ministry of Health, Institute of Endocrinology, Metabolic Disease Hospital, Tianjin Medical University, Tianjin, China, 3000703Department of pathophysiology Tianjin Medical University，Tianjin 300070，China Abstract Objectives:To investigate effects of iodide excess on Fischer rat thyroid cell line (FRTL) in early peroid. Methods: After treatment with 10-4mol/L potassium iodide (KI) for 2h, 4h, 24h, changes of mitochondrial superoxide formation were assayed by flow cytometry and fluorescence microscopy using MitoSOX. Cytochrome c (cyt c) release from mitochondria to cytoplasm was detected by immuncytochemity. Lactate dehydrogenase (LDH) activity in culture fluid was evaluated by LDH kit. Percent of dead cells were assayed by flow cytometry using propidium iodide (PI). DNA fragmentation was measured by agarose electrophoresis. Results: Mitochondrial superoxide production in FRTL cells was increased at 2h, 4h and 24h, especially at 2h. Cyt c was released from mitochondria to cytoplasm at 2h, 4h, 24h(p0.05, p0.01). LDH activity in culture medium was significantly increased at 4h, 24h(p0.01) and death cell percent was increased with treatment time. DNA ladder appeared at 24h. Conclusion: Iodide excess (10-4mol/L) can cause mitochondrial peroxide injury in FRTL cells at 2h and cell apoptosis at 24h. Keywords:iodide，thyroid gland，cells，cultured，superoxi