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目的 :探讨氧化损伤致晶体上皮细胞凋亡与白内障形成的关系。方法 :离体家兔晶体于 M199培养液中培养 ,实验组另加过氧化氢使其最后浓度为 0 .9m M,对照组不加过氧化氢。实验组与对照组均以不同时限分为 3、 6、 12、 18、 2 4、 36、 48、 6 0、 84、10 8小时 10个组。每组晶体 6～ 9只。利用白色背景下黑色方格作为判断晶体混浊度的客观指标 ,其方法是透过晶体观察方格线的清晰度并拍照。利用 TUNEL技术检测晶体上皮细胞凋亡率 ,在电子显微镜下观察细胞超微结构的改变并对凋亡细胞予以确认。结果 :在过氧化氢损伤下随着培养时间的延长 ,晶体混浊度加重 ,6小时开始轻微混浊 ,10 8小时几乎完全混浊 ;过氧化氢损伤可导致晶体上皮细胞凋亡 ,且随时间延长 ,凋亡率增高 ,3小时凋亡率为 4.45 % ,84小时凋亡率达 93.89% ,10 8小时几乎全部凋亡。结论 :过氧化氢可诱发体外培养的晶体上皮细胞凋亡 ;细胞凋亡是实验性白内障形成的细胞学基础 ;进一步探讨保护晶体免受氧化损伤及其他可致晶体上皮细胞凋亡的各种损伤因素的措施 ,就有可能防止白内障的发生
Objective: To investigate the relationship between the apoptosis of lens epithelial cells and the formation of cataract induced by oxidative damage. Methods: Rabbit crystal was cultured in M199 medium. Hydrogen peroxide was added to the experimental group to make the final concentration 0.9 M, while the control group did not add hydrogen peroxide. The experimental group and the control group were divided into 10 groups of 3, 6, 12, 18, 24, 36, 48, 60, 84, 108 hours with different time periods. Each crystal 6 to 9. The use of black squares under the white background as an objective indicator of crystal turbidity can be achieved by observing the sharpness of the grid lines through the crystal and photographing. TUNEL technique was used to detect the apoptosis rate of lens epithelial cells. The ultrastructural changes of the cells were observed under electron microscope and the apoptotic cells were confirmed. Results: With the prolongation of culture time, the turbidity of the lens increased under the injury of hydrogen peroxide. The turbidity of the lens increased slightly at 6 hours and almost completely turbid at 108 hours. Hydrogen peroxide damage led to the apoptosis of the lens epithelial cells. With time prolonging, Apoptosis rate increased 3 hours apoptosis rate was 4.45%, 84 hours apoptosis rate of 93.89%, 10 8 hours almost all apoptosis. Conclusion: Hydrogen peroxide can induce the apoptosis of cultured lens epithelial cells in vitro. Apoptosis is the cytological basis of experimental cataract formation. Further studies are conducted to investigate the effects of oxidative damage and other damage induced by crystal epithelial cells Factors of the measures, it is possible to prevent the occurrence of cataracts