Survivin基因在大鼠后发性白内障动物模型中的表达

Expression of survivin gene in animal model of posterior capsule opacification in rats

目的建立SD大鼠后发性白内障(PCO)动物模型,并检测Survivin调控基因在PCO中的表达变化,探讨Survivin对PCO生成的影响。方法取成年SD大鼠60只随机分为6组,其中10只(n=10)作为正常对照组,另50只SD大鼠于腹腔麻醉联合表面麻醉下行右眼晶状体囊外摘出术(ECLE),分为A、B、C、D、E,每10只(n=10)为1组,分别于术后即刻、3、7、14、28 d对术眼进行裂隙灯显微镜及组织病理学检查,观察PCO形成的时间、部位、发展过程及组织形态学改变,并采用逆转录聚合酶链反应(RT-PCR)方法及免疫组化方法检测Survivin基因在术后不同时间点PCO中的表达。结果 PCO在术后3 d出现,可见后囊膜皱褶,术后7、14 d后囊膜混浊明显,所有动物于术后14 d出现明显的后囊膜皱缩,可见新生晶状体纤维,术后28 d可见明显的后囊膜增厚,新生晶状体纤维填充囊袋,并且透明度下降。免疫组化和RT-PCR于正常对照组及术后即刻A组均未检测到Survivin的表达,B、C、D、E4组术后不同时段的PCO组织中可检测到不同程度Survivin的表达,其中,免疫组化显示C、D 2组表达较为明显,RT-PCR则显示C组达到表达高峰,D组表达开始下降,但较E组表达强,B组表达最弱。结论 SD大鼠可成功建立PCO动物模型并检测到Survivin调控基因的表达,Survivin的表达在PCO的形成过程中具有相应的时相性,提示Survivin与PCO的形成机制具有一定相关性,对探索PCO的基因治疗方法具有参考价值。

Objective This study is to establish animal models of posterior capsule opacification（PCO） in Sprague dawley（SD） rats and detect the expression of the survivin in PCO tissue.Methods 60 adult SD rats were divided randomly into six groups：10（n=10） in control group;extra capsular lens extraction（ECLE） were performed in the other 50 rats under abdominal cavity anesthesia combined with topical anesthesia.The operative eyes were divided into A,B,C,D,E five groups,10 in each group（n=10）,slit lamp microscope and histopathological examined were instantly after surgery and at 3 days,7 days,14 days and 28 days respectively to observe the time,location,development and histomorphological changes of PCO formation.Reverse transcription polymerase chain reaction（RT-PCR） and immunohistochemistry measurement was performed to detect the expression of the survivin in PCO at different time points postoperatively.Results 3 days after the surgery,PCO began to occur and capsular wrinkling was apparent.7 days and 14 days after the surgery,PCO became obvious.Posterior capsular wrinkling became obvious in all cases at 14 days after surgery.28 days after the surgery,the posterior capsule became obviously thicker and the regenerated lens fiber filled the capsular bag which were significantly decreased in transparency.The immunohistochemistry measurement and RT-PCR method indicated that different level of expressions of survivin could be detected in the tissue of PCO in groupB,C,D,E which at different time points postoperatively except control group and group A that instantly after surgery.The immunohistochemistry measurement indicated that expressions in group C and group D were comparatively apparent.RT-PCR method indicated that survivin reached the peak in group C and decreased in group D,lower in group E and in group B reached the bottom.Conclusion The PCO animal model can be established successfully in SD rats and the expression of survivin can be detected in the tissue of PCO in a certain similar time.The study indicated that survivin is correlate with the pathogenesis of PCO.It may provide a novel tool for the investigation of gene therapy for PCO.