Li Jian-Ming, Zhou Hong, Cai Qian, Xiao Guang-Xia
Institute of Burn Research, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
World J Gastroenterol. 2003 Mar;9(3):562-7. doi: 10.3748/wjg.v9.i3.562.
To study the role of mitochondrial dysfunction in hydrogen peroxide-induced apoptosis of intestinal epithelial cells.
Hydrogen peroxide-induced apoptosis of human intestinal epithelial cell line SW-480 was established. Cell apoptosis was determined by Annexin-V and PI double-stained flow cytometry and DNA gel electrophoresis. Morphological changes were examined with light and electron microscopy. For other observations, mitochondrial function, cytochrome c release, mitochondrial translocation and membrane potential were determined simultaneously.
Percentage of apoptotic cells induced with 400 micro mol/L hydrogen peroxide increased significantly at l h or 3 h after stimulation and recovered rapidly. Meanwhile percentage of apoptotic cells induced with 4 mmol/L hydrogen peroxide increased with time. In accordance with these changes, we observed decreased mitochondrial function in 400 micro mol/L H2O2-stimualted cells at 1 h or 3 h and in 4 mmol/L H2O2-stimualted cells at times examined. Correspondingly, swelling cristae and vacuole-like mitochondria were noted. Release of cytochrome c, decreased mitochondrial membrane potential and mitochondrial translocation were also found to be the early signs of apoptosis.
Dysfunctional mitochondria play a role in the apoptosis of SW-480 cell line induced by hydrogen peroxide.
研究线粒体功能障碍在过氧化氢诱导的肠上皮细胞凋亡中的作用。
建立过氧化氢诱导人肠上皮细胞系SW - 480凋亡的模型。采用Annexin - V和PI双染流式细胞术及DNA凝胶电泳检测细胞凋亡。用光镜和电镜观察形态学变化。同时测定线粒体功能、细胞色素c释放、线粒体转位及膜电位等指标。
400 μmol/L过氧化氢诱导的凋亡细胞百分比在刺激后1小时或3小时显著增加，随后迅速恢复。同时，4 mmol/L过氧化氢诱导的凋亡细胞百分比随时间增加。与这些变化一致，我们观察到在1小时或3小时时400 μmol/L H₂O₂刺激的细胞以及在各检测时间点4 mmol/L H₂O₂刺激的细胞中线粒体功能下降。相应地，观察到嵴肿胀和线粒体呈空泡样。细胞色素c释放、线粒体膜电位降低和线粒体转位也被发现是凋亡的早期迹象。
功能失调的线粒体在过氧化氢诱导的SW - 480细胞系凋亡中起作用。
