去整合素Echistatin抑制糖尿病兔远期后发性白内障

Different concentrations of disintegrin Echistatin in inhibition of long-term posterior capsule opacification in diabetic rabbit

目的观察并探讨不同浓度去整合素(Echistatin,Ecs)对糖尿病兔远期后发性白内障(posterior capsular opacification,PCO)发生过程中晶状体上皮细胞(lens epithelial cells,LEC)增殖的抑制作用,以及其在眼内的安全性。方法建立糖尿病兔模型,随机分为对照组(A组,术毕前房注入蒸馏水0.2 mL);实验组:B组、C组、D组(术毕前房分别注入5.0×10-3μg·L-1、7.5×10-3μg·L-1、10.0×10-3μg·L-1Ecs),术后裂隙灯下观察角膜、前房及PCO情况。术后6周取材,常规石蜡切片,HE染色。用免疫组织化学法检测PCO发生过程中LEC增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达,TUNEL法检测角膜内皮细胞、视网膜神经节细胞层及内核层细胞的凋亡情况。结果 HE染色结果显示,术后6周各组角膜内皮细胞排列整齐、未见水肿,视网膜内界膜与各层组织结构完整,未见药物引起明显的组织损伤反应。各组间PCO分级差异有统计学意义(P=0.011)。免疫组织化学法检测各组后囊膜PCNA表达结果显示,A组、B组、C组、D组的平均光密度值分别为0.499±0.043、0.484±0.055、0.422±0.012、0.340±0.031,B组后囊膜上PCNA表达与A组差异无统计学意义(P>0.05),余组间两两比较差异均有统计学意义(均为P<0.05)。TUNEL法检测各组角膜内皮细胞、视网膜神经节细胞层及内核层细胞凋亡发现,各组间细胞凋亡比较均未见明显差异(均为P>0.05)。结论 10.0×10-3μg·L-1Ecs对糖尿病兔远期PCO具有明显抑制作用,且对眼内相邻及重要组织无明显毒性作用,因此可选用此浓度进行后续研究。

Objective To observe the inhibitive effects of different concentrations of disintegrin Echistatin （ Ecs ） on lens epithelial cell （ LEC ） proliferation during long-term posterior capsular opacification（PCO）development,and discuss the safe concentration in the eyes of rabbits. Methods Diabetic rabbit models were established and randomly divided into control group （group A, surgery anterior chamber were injected 0.2 mL of distilled water）and experimental groups（ group B, C and D, surgery anterior chamber were injected 0.2 mL Ecs of 5.0 × 10^-3 μg · L^-1 ,7.5 × 10^-3 μg · L^-1 , 10.0 × 10^-3 μg · L^-1, respectively）. After injection, cornea, anterior chamber and PCO were observed under slit lamp. At postoperative 6 weeks, HE staining was performed, proliferating cell nuclear antigen（PCNA） expression in LEC during PCO development was observed by immunohistochemistry, and TUNEL assay was used to detect the apoptosis of corneal endothelial cells, retinal inner nuclear layer cells and ganglion cells. Resuits HE staining showed that the corneal endothelial cells arranged regularly with no edema, the structure of retinal internal limiting membrane and each layer of tissue were unchanged,no significant drug-induced damage tissue was appeared in each group at postoperative 5 weeks. There was statistical difference in PCO grading among groups （P =0. 011 ）. Immunohistochemistry staining showed that the average optical density of PCNA expression in posterior capsule of group A, B, C and D were 0. 499 ±0. 043, 0.484 ±0. 055,0.422 ±0.012 and 0. 340 ±0.031 ,respectively,there were statistical differences between each two groups except for group A and group B（ all P 〈 0.05 ）. TUNEL method show that there was no statistical difference in the apoptosis of corneal endothelial cells, retinal hmer nuclear layer cells and ganglion cells between each two groups （ all P 〉 0.05 ）. Conclusion Ecs with 10.0 × 10^-3 μg · L^-1 can not only obviously inhibit the development of long-term PCO in diabetic rabbits,but has no obvious toxic effects on intraocular adjacent tissues,so 10.0× 10^-3 μg · L^-1 Ecs can be used in further study.