基质金属蛋白酶抑制剂GM6001抑制大鼠脉络膜新生血管形成的初步研究

摘要

Objective To explore the efficiency of GM6001 for the inhibition of choroidal neovascularization. Methods Experimental study. Twenty-four Brown Norvy ( BN ) rats after photocoagulation were randomly divided into 3 groups as GM6001 group,DMSO group and CONT group.GM6001 (0.2 ml of 0.1％ suspension) was injected retrobulbarly for GM6001 group and0.2 ml DMSO was injected for DMSO group on days 1,3,6,9,and 12 after photocoagulation.No injection was performed in the CONT group.Fundus fluorescence angiography,histopathology,immunohistochemistry and quantitative analysis of choroidal neovascularization ( CNV ) were performed 3 weeks after photocoagulation. One-way ANOVA was used in conjunction with SNK-t test to assess statistical significance within groups. Resufts The fluorescein leakage appeared in all three groups; but fluorescein leakage of GM6001 group( 74.56 ± 2.33 ) was less than that of DMSO group ( 119.57 ± 1.15 ) and CONT group ( 122.36 ± 2.38 ) ( F =403.23,P =0.001 ;LSD-t test,all P value ＜0.01 ),whereas fluoresecin leakage of DMSO group was similar to that in the CONT group.The retinal and choroidal capillaries in the CONT group were damaged and disordered;a great deal of CNV and migration and proliferation of retinal pigment epithelium cells,fibrocytes and collagen fibers were discovered.Pathological changes in DMSO group were similar to those in the CONT group.There were a small quantity of retinal pigment epithelium cells,fibrocytes and CNV in GM6001 group.Although the immunohistochemical staining for CD105 displayed positive results in all three groups,positive staining of GM6001 group ( 19.85 ± 1.59 ) was significantly less than that of the CONT group (38.02 ± 2.57 ) and DMSO group (39.02 ± 3.12 ) (F =55.57,P =0.001 ; LSD-t test,all P value ＜ 0.01 ).Positive staining of CD105 in the CONT group was similar to that of DMSO group (P ＞ 0.05 ).The size of CNV in GM6001 group ( 15.35 ±0.77) was significantly less than that of CONT group( 28.38 ± 1.60 ) and DMSO group(28.74 ± 1.19) ( F =114.85,P =0.001 ; LSD-t test,all P value ＜ 0.01 ),There was no statistical difference for the size of CNV between CONT group and DMSO group (P ＞ 0.05). Conclusion GM6001 effectively inhibits CVS induced by krypton laser photocoagulation.%目的 探讨基质金属蛋白酶抑制剂GM6001对大鼠脉络膜新生血管(CNV)的抑制效果.方法 实验研究.24只BN大鼠氪激光光凝视网膜后随机分为GM6001组、二甲基亚砜( DMSO)组和空白对照(CONT)组,每组8只.GM6001组和DMSO组分别于光凝后即刻、3、6、9、12 d球后注射0.1％ GM6001混悬液0.2 ml和DMSO 0.2 ml,CONT组不做处理.光凝后3周行荧光素眼底血管造影( FFA)、组织病理学检查、CD105免疫组织化学和脉络膜铺片CNV定量检查.多组间数据的比较采用方差分析,其均数的两两比较采用LSD-t检验.结果 GM6001组、DMSO组和CONT组大鼠视网膜光凝区均可见荧光素渗漏；GM6001组(74.56±2.33)荧光素渗漏范围明显小于DMSO组(119.57±1.15)和CONT组(122.36±2.38)(F=403.23,P=0.001；LSD-t检验,P值均＜0.01)；DMSO组与CONT组的荧光素渗漏范围相似.光镜下观察CONT组光凝区可见大量新生血管、分层增殖迁移的视网膜色素上皮细胞、成纤维细胞和胶原纤维；DMSO组与CONT组观察结果相似；GM6001组光凝区增殖的视网膜色素上皮细胞、成纤维细胞和新生血管较少.3组光凝区均可见CD105阳性表达；GM6001组(19.85±1.59) CD105的表达水平明显低于CONT组(38.02±2.57)和DMSO组(39.02±3.12)(F=55.57,P=0.001；LSD-t检验,P值均＜0.01)；而CONT组与DMSO组的差异无统计学意义(P＞0.05).GM6001组CNV面积(15.35±0.77)与CONT组(28.38±1.60)和DMSO组(28.74±1.19)比较,差异有统计学意义(F=114.85,p=0.001；LSD-t检验,P值均＜0.01).结论 GM6001可以显著抑制氪激光诱发的CNV的形成.