去整合素kistrin对兔晶状体上皮细胞凋亡及连接蛋白43表达的影响

Effect of disintegrin kistrin on apoptosis of rabbit lens epithelial cells and expression of connexin43

目的研究去整合素kistrin对兔晶状体上皮细胞(lens epithelial cells,LEC)的诱导凋亡作用,初步探讨这种凋亡作用与细胞连接蛋白43(connexin43,Cx43)表达变化的联系。方法取12只新西兰大白兔,随机分为实验组与对照组,每组6只,行右眼透明晶状体囊外摘除术,建立兔后囊膜混浊模型。术毕实验组囊袋内注入浓度为80μg.L-1的kistrin0.2mL,对照组注入等量生理盐水。术后1d、3d、5d、7d、14d在裂隙灯下观察实验动物眼部情况。术后14d处死动物,摘取眼球,进行常规石蜡切片,HE染色;TUNEL法检测LEC凋亡情况,计算出凋亡指数;间接免疫荧光法检测LEC中Cx43表达情况。结果术后实验组与对照组角膜眼前段情况无明显差异。HE染色显示对照组瞳孔区后囊单层细胞黏附,而实验组后囊则保持光滑。TUNEL法检测发现实验组LEC凋亡指数为(48.82±9.99)%,明显高于对照组的(12.36±7.53)%,2组比较差异有统计学意义(P<0.01);实验组LEC中Cx43荧光阳性表达面积比为(0.03±0.02)%,对照组为(0.14±0.06)%,实验组明显低于对照组,差异有统计学意义(P<0.05)。结论去整合素kistrin在兔活体内可引起LEC间连接发生变化且可诱导LEC凋亡。

Objective To investigate the apoptosis-induced effect of disintegrin kistrin on rabbit lens epithelial cells（LEC）,and explore the relation between the apoptosis and the change of connexin43（Cx43） expression.Methods Twelve New Zealand rabbits were randomly divided into experimental group and control group,6 cases in each group,and the extracapsular lens extraction was performed on the right eye of each rabbit to establish the model of posterior capsule opacification（PCO）.The capsular bag of lens were injected with 80 μg·L-1 of kistrin 0.2 mL in the experimental group and the normal saline 0.2 mL in the control group just after the operation.All eyes were examined by slit-lamp microscopy at 1 day,3 days,5 days,7 days and 14 days after the operation.The eyeballs were extracted,paraffin embedded and then stained by HE at 14 days after the operation.The apoptosis of LEC was detected by TUNEL,the apoptosis index of LEC was calculated,and the expression of Cx43 was detected by indirect immunofluorescence.Results There was no significant difference in the conditions of anterior segment between the two groups.HE staining showed that the single layer cells adhered in the posterior capsule around the pupil in the control group,but the posterior capsule in the experimental group was smooth.The apoptosis index of LEC in the experimental group,which was（48.82±9.99）%,was higher than that in the control group,which was（12.36±7.53）%,there was significant difference（P〈0.01）.The ratio of Cx43 positive fluorescent area in LEC in the experimental group,which was（0.03±0.02）%,was less than that in the control group,which was（0.14±0.06）%,there was statistical difference（P〈0.05）.Conclusion The disintegrin kistrin can change the connections between LEC and induce its apoptosis in vivo.