米诺环素对糖基化终末产物所致大鼠视网膜微血管渗漏的作用及其机制研究  

Effects of minocycline on leakage of retinal microvessels induced by glycation end products and its mechanism

作　　者：陈永东[1,2] 许迅[1] 吴强[2] 郑志[1] 朱冬青[1] 顾青[1] 

机构地区：[1]上海交通大学附属上海市第一人民医院眼科,博士研究生200080 [2]上海交通大学附属上海市第六人民医院眼科,200233

出　　处：《眼科研究》2010年第12期1105-1109,共5页Chinese Ophthalmic Research

摘　　要：目的研究基质金属蛋白酶-9(MMP-9)在糖基化终末产物(AGEs)所致血-视网膜屏障损害中的作用,评估米诺环素对AGEs所致血-视网膜屏障损害的治疗作用。方法 36只SD大鼠用随机数字表法分为牛血清白蛋白(BSA)组、AGEs-BSA组和AGEs-BSA+米诺环素组。BSA组和AGEs-BSA组分别自尾静脉注入BSA和AGEs-BSA,每日1次,每次40mg/kg,AGEs-BSA+米诺环素组在每日注射AGEs-BSA的同时隔日腹腔注射米诺环素45mg/kg。14d后,每组取6只大鼠用伊文思蓝(EB)法观察并比较各组视网膜血管的通透性,其余大鼠处死后,采用RT-PCR及Westernblot法观察视网膜MMP-9的表达。结果 AGEs-BSA组MMP-9mRNA和蛋白水平均较BSA组升高,差异有统计学意义(P<0.01),同时,视网膜微血管渗透性也显著升高(P<0.01)。而在AGEs-BSA+米诺环素组大鼠视网膜基因和蛋白水平的MMP-9均被抑制(P<0.01),同时视网膜微血管渗漏被部分逆转(P<0.01)。视网膜微血管的渗透性与MMP-9蛋白的表达呈正相关(r=0.891,P=0.000)。结论 AGEs所致的视网膜微血管渗漏与MMP-9的高表达有密切关系,AGEs的作用至少部分是通过MMP-9实现的。米诺环素能部分逆转AGEs-MMP-9途径所致的血管渗漏。Background Advanced glycation endproducts（AGEs）is an important etiological factor of diabetic retinopathy,and they result in the damage of vascular endothelial cells and apoptosis of perithelial cells.However,it is unclear that how they influence the retina capillary permeability.ObjectivePresent study was to study the role of matrix metalloproteinase 9（MMP-9）in blood-retinal barrier damage induced by AGEs and explore the therapeutic effects of minocycline,a MMPs inhibitor.MethodsThirty-six SPF Sprague-Dawley rats were randomly assigned to 3 groups and 12 for each group.Bovine serum albumin（BSA）（40mg/d）was injected via caudal vein in the rats of the control group.AGEs＋BSA was used at the same way in 12 rats to establish the blood-retinal barrier damage models.In minocycline group,minocycline（45mg/kg）was intraperitoneally injected at the same time of injection of AGEs＋BSA at the interval of 1 day in 12 model rats.After 2 weeks of injection,the retinas of experimental animals were obtained.The levels of MMP-9 protein and mRNA in retina were detected by Western blotting and semi-quantitative real time reverse transcription-polymerase chain reaction（RT-PCR）,respectively.The permeability of retinal vessels was bio-assayed using the Evans blue method.The correlation between the vessel permeability and the MMP-9 protein levels was analyzed.ResultsThe gray-scale value of MMP-9 mRNA was 0.86±0.045,1.12±0.053,0.91±0.07 in BSA group,AGEs-BSA group and minocycline group,showing a significant difference among three groups（F=37.090,P=0.000）.Compared with the BSA group,the MMP-9 mRNA level in the retina was significantly increased in AGEs-BSA group（P0.05）,and that in minocycline group was considerably lower than AGEs-BSA group（P0.05）.The gray-scale value of MMP-9 protein was 0.204±0.033,0.512±0.083 and 0.371±0.056 in BSA group,AGEs-BSA group and minocycline group respectively with a significant difference among them（F=38.470,P=0.000）,and expression of MMP-9 protein was upregulat

关 键 词：米诺环素 基质金属蛋白酶-9 糖基化终末产物 糖尿病视网膜病变 血-视网膜屏障 

分 类 号：R774.1[医药卫生—眼科]