摘要
目的 观察过氧化氢对体外培养肠上皮细胞系SW 480线粒体的损伤作用。方法 以不同浓度的过氧化氢处理人肠上皮细胞株 (SW 480 ) ,观察细胞形态学和线粒体超微结构的改变。以MTT法测定线粒体整体功能的变化。应用DNA凝胶电泳和Annexin V及PI双标流式细胞术观察细胞凋亡的发生。应用免疫组织化学法检测细胞色素c的释放。结果 40 0 μmol L过氧化氢在 3h可引起线粒体整体功能的明显改变 ,但以后逐渐恢复。 4mmol L过氧化氢可引起线粒体整体功能不可逆的进行性下降 ;可引起线粒体空化 ,嵴肿胀。随着过氧化氢浓度的增加 ,线粒体的空化更加明显 ;可引起细胞发生凋亡。过氧化氢处理早期即出现细胞色素c的释放。结论 过氧化氢可引起肠上皮细胞线粒体明显损伤 。
Objective To study the role of hydrogen peroxide in the pathogenesis of the mitochondria in cultured intestinal epithelial cell line SW 480. Methods Cultured intestinal epithelial cells were treated with hydrogen peroxide at different concentrations. The cell morphology and the mitochondial ultrastructure were observed with invert and transmission electron microscopy respectively. MTT assay was used to measure mitochondrial function. The apoptosis of SW 480 cells was determined with DNA gel electrophoresis and Annexin V and PI double stained flow cytometry. The Release of cytochrome c from the mitochondria was detected with immunohistochemistry. Results Hydrogen peroxide of 400 μmol/L gradually caused reversible changes on mitochodial functions of the cells, but for the cells treated with 4 mmol/L hydrogen peroxide, the changes were irreversible and became more serious with the increase of agent dose. The mitochondria were vacuolated and their cristae were swollen in a dose dependent manner under microscope. Hydrogen peroxide led to the apoptosis of SW 480 cells and the release of cytochrome c in the early stage of treatment. Conclusion Hydrogen peroxide causes distinct injuries on the intestinal epithelial cell mitochondria, whose structure and functions are closely correlated with cell apoptosis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2002年第2期142-145,共4页
Journal of Third Military Medical University
基金
国家重点基础研究发展规划项目 ("973"项目 ) (G19990 54 2 0 2 )
