缺血再灌注大鼠视网膜微血管基膜细胞外基质与视网膜细胞线粒体钙含量的变化  

Changes of extracellular matrix of microvascular basement membrane and mitochondrial calcium concentration in retina of rats during ischemia and reperfusion

作　　者：范定怀[1] 陈少强[2] 黄焱[2] 张更[2] 

机构地区：[1]福建省莆田学院基础医学部,福建省莆田市351100 [2]福建医科大学解剖学与组织胚胎学系,福建省福州市350004

出　　处：《中国临床康复》2005年第30期144-146,i0005,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:采用前房灌注加压法制造视网膜缺血再灌注模型,用抗Ⅳ型胶原、层粘连蛋白和纤维粘连蛋白抗体对大鼠视网膜微血管基膜的细胞外基质进行免疫组化反应,观察视网膜微血管基膜的细胞外基质和视网膜细胞线粒体钙含量的变化。方法:实验在福建医科大学临床应用解剖学研究室完成。90只大鼠随机分为正常对照组和缺血1h再灌注1h组、缺血1h再灌注2h组、缺血2h再灌注1h组、缺血2h再灌注2h组,每组18只。缺血再灌注各组采用提高眼压法造成视网膜缺血后,恢复眼压形成血流再灌注,每组取8只大鼠视网膜组织用抗Ⅳ型胶原、层粘连蛋白和纤维粘连蛋白抗体标记,测定阳性染色平均单位面积。每组取10只大鼠测定视网膜细胞线粒体钙浓度的变化。结果:90只大鼠均进入结果分析。①视网膜微血管基膜的Ⅳ型胶原、层粘连蛋白和纤维粘连蛋白阳性染色平均单位面积:缺血再灌注各组比正常对照组小(P<0.01)。②视网膜微血管基膜阳性产物呈不连续线状的数量:缺血再灌注各组比正常对照组多(P<0.01)。③视网膜细胞线粒体钙含量:缺血1h再灌注1,2h组,缺血2h再灌注1,2h组均高于正常对照组[(21.82±1.97)μg/g,(40.45±3.92)μg/g,(43.68±4.01)μg/g,(68.76±5.37)μg/g,(13.37±1.23)μg/g,(F=6.634～8.987,P<0.01)]。随缺血再灌注的延长,大鼠视网膜线粒体钙含量逐渐增高。结论:缺血再灌注可引起视网膜细胞线粒体钙含量升高,并可能激活细胞外的蛋白酶,降解微血管基膜细胞外基质的主要成分-Ⅳ型胶原、层粘连蛋白和纤维粘连蛋白,破坏血视网膜屏障,导致视网膜组织结构和功能损伤。AIM： Retina ischemia-reperfusion （RIR） model was prepared with anterior chamber perfusion with pressure. Immunohistochemical reaction was performed on extracellular matrix of retinal microvascular basement membrane of the rats with antidies against type Ⅳcollagen, laminin and fibronectin . To observe the change of extracellular matrix of retinal mierovascular basement membrane and concentration of mitrochondrialcalcium. METHODS： The experiment was conducted in the Institute of Clinical Application Anatomy, Fujian Medical University. Totally 90 rats were randomly divided into normal control group, ischemia 1 hour and reperfusion 1 hour group, ischemia lhour and reperfusion 2 hours group, ischemia 2 hours and reperfusion 1 hour group, ischemia 2 hours and reperfusion 2 hours group, with 18 rats in each group. The intraocular pressure （IOP） was raised to occlcde retinal blood vessels to form retinal ischemia, and then IOP was lowered normal to allow reperfusion. Retinal tissue of 8 rats in each group was labeled with antibodies against type Ⅳ collagen, laminin and fibronectin , and the average unit area of positive staining was measured. Retinal tissue of 10 rats in each group was measured for concentration of mitrochondrial calcium. RESULTS： Totally 90 rats entered the stage of result analysis.① average unit area of positive staining of type Ⅳ collagen, laminin and fibronectin of retinal microvascular basement membrane in the ischemia-reperfusion groups was smaller than that in the control group （P 〈 0.01）. ② Discontinuous line positive product of retinal microvaseular basement membrane was more in the isehemia-reperfusion group than that in the normal control group （P 〈 0.01）.③Concentration of mitochondrial calcium of retina was all higher in the ischemia 1 hour and reperfusion 1 hour group, ischemia 1 hour and reperfusion 2 hours group, ischemra 2 hours and repeffusion 1 hour, ischemia 2 hours and repeffusion 2 hours group than that in the normal control group [（21.82±1.97

关 键 词：缺血 再灌注 视网膜 线粒体 钙 细胞外基质 大鼠 

分 类 号：R774[医药卫生—眼科]