纤溶酶K区缺失突变体玻璃体腔注射对光化学诱导大鼠视网膜分支静脉阻塞的溶栓作用  

Thrombolysis of plasmin-AK on photochemicaily induced branch retinal vein occlusion after intravitreaiinjection in rats

作　　者：陈武[1] 莫炜[3] 黄欣[4] 欧阳艳玲[4] 宋后燕[3] 张志云[2] 

机构地区：[1]湖北医药学院附属东风医院检验科,湖北省十堰市442001 [2]湖北医药学院附属东风医院外科,湖北省十堰市442001 [3]复旦大学教育部代谢与分子医学重点实验室,上海200032 [4]复旦大学附属眼耳鼻喉医院,上海200031

出　　处：《中华实验眼科杂志》2016年第5期408-413,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：湖北省教育厅资助项目（B2013105）

摘　　要：背景视网膜静脉阻塞是常见的视网膜血管性疾病，目前溶栓和抗凝疗法是重要的治疗手段。然而，系统溶栓疗法效率较低，且易增加出血风险。目的观察玻璃体腔注射纤溶酶K区缺失突变体（PLM—AK）对光化学诱导的大鼠视网膜分支静脉阻塞（BRVO）的治疗作用。方法用SD大鼠尾静脉注射孟加拉玫瑰红溶液40mg／kg，然后用氩激光照射视网膜静脉法建立SD大鼠BRVO模型，采用随机数字表法将造模成功的40只大鼠随机分为平衡盐溶液（BSS）组、0．01U（商品单位）PLM—AK组、0．02UPLM—AK组和0．03UPLM—AK组，每组10只。造模并避光饲养大鼠12h后于大鼠玻璃体腔内分别注射BSS和0．01、0．02或0．03UPLM—AK10μl，各组大鼠于注射后3d行间接检眼镜、荧光素眼底血管造影（FFA）检查。用过量麻醉法处死SD大鼠并制备视网膜铺片和眼球壁切片，采用苏木精一伊红染色法观察大鼠球壁的形态学变化；采用免疫荧光法检测大鼠球壁组织中人纤连蛋白（FN）和层黏连蛋白（LN）的表达；透射电子显微镜下观察大鼠视网膜的超微结构改变。结果玻璃体腔内药物注射后3d，FFA显示BSS组及0．01、0．02或0．03UPLM—AK组视网膜分支静脉再通达2支以上的大鼠数量分别为0、3、6和8只，组间总体比较差异有统计学意义（x2=9．635，P=0．022），其中0．01UPLM—AK组再通血管的大鼠数量与BSS组比较，差异无统计学意义（Z=-1．558，P=0．119），而0．03UPLM—AK组再通血管的大鼠数量明显多于0．01UPLM—AK组，差异有统计学意义（Z=-2．762，P=0．006）。玻璃体腔药物注射后3d，BSS组大鼠视网膜静脉内可见血栓形成，视网膜铺片可见新生血管；而0．03UPLM-AK组可见大鼠玻璃体后脱离，视网膜铺片未见新生血管形成。BSS组可见FN主要表达于内界膜（ILM）层、感光细胞层（PCL）、外界膜（OLM）层、脉络膜和巩Background Retinal vein occlusion is a common retinal vascular diseases. Thromblysis and anticoagulation therapies are main approaches. However, systemic thrombolysis is relatively inefficient, and it often enhances the risk of hemorrhage. Objective This study was to investigate the therapeutic effects of PLM-AK,a kringle deficiency mutant of plasmin, on photochemically induced branch retinal vein occlusion （BRVO） after intravitreal injection. Methods BRVO models were established by the combination of caudal vein injection of RoseBengal with argon laser radiation of periphery area of retinal veins in SD rats. Forty model rats were randomized into balance salt solution （BSS） group and O. 01 U,0. 02 U,0. 03 U PLM-AK group, and 10 μl corresponding drug was intravtreally injected 12 hours after modeling. Ophthalmoscopy and fundus fluorescein angiography （FFA） were performed to observe the change of retinal veins. The animals were sacrificed 3 days after intravitreal injection, and hematoxylin and eosin staining was used for the histopathological and ultrastructural examination of retinas. The retina of the rats was isolated for the stretched preparation of retina. The expressions of fibronectin （FN） and laminin （LN） in eyeball wall were assayed by immunofluorescence technology. The use and care of the animals complied with Statement of the Association for Research in Vision and Ophthalmology. Results The revascularization of over 2 retinal veins was found in 0,3,6 and 8 rats in the BBS group and 0.01 U ,0.02 U ,0.03 U PLM-AK group 3 days after intravitreal injection, respectively, showing a significant difference among the groups （X2 = 9. 635 ,P = 0. 022）, and the rat number with revascularization in 0.01 U PLM-AK group was not significantly different from that in BSS group （Z=-I. 558,P = 0. 119 ）, but the difference between 0. 03 U PLM-AK group and 0. 01 U PLM-AK group was significant （Z=-2. 762 ,P-0. 006）. In the third day after intravitreal injection, retinal vein thrombus were fou

关 键 词：视网膜静脉阻塞/治疗 纤维蛋白溶解剂/用法及用量 玻璃体注射 溶栓疗法/方法 动物模型 SD大鼠 

分 类 号：R774[医药卫生—眼科]