摘要
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 。
Objective:To investigate the relationship between lens epithelial cell apoptosis induced by H 2O 2and cataract formation.Methods:Vitro rabbits lenses were incubated with M199 involving 0 9mM H 2O 2(experimental groups)and exclusive of H 2O 2(control groups).Experimental and control groups were divided into ten groups which were incubated for 3h,6h,12h,18h,24h,36h,48h,60h,84h,108h.Each group included 6～9 lenses.Lenses opacification were detected with black squared paper and took pictures.Apoptotic lens epithelial cells was assayed using the TUNEL technique.Transmission electron microscopy verified the occurrence of programmed cell death.Result:①the longer time incubate With H 2O 2 treatment,the more opaque of lens was found.Lenses opacification were found at 6 hours after treatment with H 2O 2,and the lenses became almost completely opaque in about 108 hours after treatment with H 2O 2.②During incubation of lenses with H 2O 2 treatment,the percentage of apoptotic cells gradually rose as the incubation time increased.About 4 45% of the epithelial cell nuclei were TUNEL positive at 3 hours after treatment with H 2O 2.After 84 hours incubation,more than 93 89% of the lens epithelial cells underwent apoptosis,and almost all the lens epithelial cells became apoptosis after 108 hours treatment with H 2O 2.③H 2O 2induced lens epithelial cell apoptosis precedes cataract formation.Conclusion:H 2O 2 induced lens epithelial cell apoptosis was a cellular basis for cataract development induced by H 2O 2.
出处
《中国实用眼科杂志》
CSCD
2000年第11期669-672,T001,T002,共6页
Chinese Journal of Practical Ophthalmology
