毛果芸香碱滴眼对兔眼角膜内皮细胞凋亡作用的影响.pdfVIP

第45卷第12期 山 东 大 学 学 报 (医 学 版) Vo1．45 No．12 2OO7年 12月 JOURNAL OF SHANDONG UNIVERSITY(HEALTH SCIENCES) Dec．2唧 文章编号：1671—7554(2007)12—1271—04 毛果芸香碱滴眼对兔眼角膜内皮 细胞凋亡作用的影响 陶远，王红，乔智 (山东大学齐鲁医院眼科，济南250012) 摘要：目的 探讨长期应用毛果芸香碱对兔眼角膜内皮细胞有无影响及影响方式。方法 用2％的毛 果芸香碱给兔滴眼1个月和3个月后，取下兔眼球迅速固定，分离角膜内皮组织，用蛋白印迹(Western．blot) 法检测兔眼角膜内皮细胞中抗凋亡基因bc1．2蛋白及凋亡基因bax蛋白的表达，流式细胞仪检测角膜内皮细 胞的凋亡率，透射电镜下观察各组角膜内皮细胞的超微结构变化。结果 应用毛果芸香碱1个月及3个月 后，均呈现兔眼角膜内皮细胞凋亡基因bax蛋白表达增多，抗凋亡基因bc1．2蛋白表达减少，3个月组兔眼角 膜内皮细胞凋亡率高于对照组(P0．05)，透射电镜下观察可见，1个月及3个月组细胞内空泡形成，线粒体 肿胀，内质网扩张，细胞核固缩，染色质固缩周边化等典型凋亡改变。结论 长期应用2％的毛果芸香碱可 以诱导兔眼角膜内皮细胞发生凋亡。 关键词：细胞凋亡；角膜内皮；毛果芸香碱；bax蛋白；bc1．2蛋白 中图分类号：R775．9 文献标志码：A Effect of pilocarpine on apoptosis of corneal endothelium in rabbits TAO Yuan，WANG Hong，QIAO Zhi (Department of Ophthalmology，Qilu Hospital of Shandong University，Jinan 250012，China) AbStract：Objective To investigate the effects and mechanisms of pilocarpine on corneal endothelium in rabbits． Methods 20 rabbits in the experiment group were subjected to 2％pilocarpine 3 times a day for 1 month or 3 months． The expressions of bcl-2 and bax protein were determined by Westernblot，the apoptosis rate of rabbit corneal endothelium cells was determined by flow cytometry，and the morphological changes of the corneal endothelium cells were determined by transmission electron microscopy．Results The expression of bax protein in the experimental group Was higher thah that in the control group，however the expression of bcl-2 protein Was lower than that in the control group．The apoptosis rate of the rabbit come,a]endothelium cells using pilocarpine for 3 months Was significantly higher than that of the control group(P0．05)．Also transmission electronic microscopy revealed there were morphological ch