改良大鼠后囊膜混浊模型的建立及晶状体上皮细胞的变化

目的:改良法建立大鼠后发性白内障动物模型并观察LEC在PCO过程中的动态变化。方法:SD大鼠50只在连续环形撕囊、水分离后行晶状体囊外摘除术(ECLE),娩核后使用无菌空气恢复前房。分别于术后0,3,7,14及28d对术眼进行裂隙灯检查并处死动物,摘除眼球行光镜观察,免疫组化检测LEC的α-SMA表达。结果:100%术眼后囊膜存在,84%的鼠眼可用于检测。术后0h于前囊下和赤道部的内表面观察到LEC。PCO在术后3d出现,出现囊膜皱缩,整个晶体囊膜出现纺锤形细胞。术后7d时后囊膜明显混浊,见较多纺锤形细胞分布。所有动物于术后14d出现明显后囊膜皱缩,可见新生晶体纤维。术后28d见明显后囊膜增厚,新生晶体纤维填充囊袋,后囊膜未见细胞。LEC形态及分布恢复到术后0h状态。免疫组化检测见术后3d时α-SMA阳性表达。结论:改良法成功建立大鼠PCO模型,LEC在PCO形成中出现形态和分布上的动态变化。其将为在分子水平上探索PCO的发病机制及防治方法提供合适研究载体。

Establishment of an improved model of posterior capsule opacification in rats and dynamic observation of LEC

AIM:To establish an animal model of posterior capsule opacification (PCO) in rats by an improved surgical procedure and to observe the dynamic changes of lens epithelial cells (LEC) during the development of PCO.METHODS: An extracapsular lens extraction (ECLE), after continuous curvilinear capsulorrhexis and hydrodi-ssection, was performed in 50SD rats. Bacteria-free air was used to fill and maintain the anterior chamber after the lens was removed. Animals were observed by slit-lamp microscope at 0 hour and 3, 7, 14 and 28 days after the surgery, and were killed at these time points. The eyeballs were enucleated for light microscopy. The transdifferentiation of the LEC was indicated by the detection of alpha-smooth muscle actin (α-SMA) through immunohistochemistry.RESULTS: All of the posterior capsules were integrate after ECLE, and 84% operated eyes recovered well and could be used to detect. The lens epithelial cells (LEC) could be seen in the inner surface of equator and under the anterior capsule. At 3 days after the surgery, PCO began to occur, and spindle shaped cells were present throughout the lens capsule. Capsular wrinkling was apparent. At 7 days the PCO was obvious and several fibrocytes migrated to the posterior capsule. Capsular wrinkling could be found at 14 days after the surgery in all the animals, and lens fiber could be observed under the light microscope. At 28 days after the surgery the thickness of posterior capsule increased obviously and the regenerated lens fiber filled the capsular bag. Cells were no longer found in the posterior capsule. The form and distribution of LECs recovered to those of 0 hour after the surgery. At 3 days after surgery α-SMA was found positive expression in the LEC of posterior capsule.CONCLUSION: The PCO animal model can be established successfully in rats by an improved surgical procedure. The dynamic changes of LEC can be found during the development of PCO. It is a proper model for the investigation of the pathogenesis, prevention and treatment of PCO on molecular level.