GM6001抑制视网膜新生血管形成VEGF和MMP2的表达

Inhibition of GM6001 on retinal neovascularization in neonatal rats with retinopathy

目的:探讨基质金属蛋白酶抑制剂GM6001对鼠视网膜新生血管形成的抑制作用及机制。方法:采用高氧使鼠视网膜血管阻塞建立早产儿视网膜病变模型后,将鼠分为两组,玻璃体腔内注射75μmol/L的GM60010.5μL,另一组不治疗。5d后处死,行视网膜HE染色计数矢状面5μm视网膜切片中突破内界膜的血管内皮细胞核数,和血管内皮生长因子(VEGF)和基质金属蛋白酶-2(MMP-2)免疫组织化学染色,以及视网膜铺片。同时设立正常对照组。结果:GM6001治疗组视网膜发生新生血管的眼数较高氧对照组少,但较正常对照组多。GM6001治疗组、高氧对照组、正常对照组平均每个切面突破内界膜的血管内皮细胞核数为2.38±0.90,21.25±1.12和1.23±0.46,高氧对照组与正常对照组具有显著统计学差异(P<0.01),GM6001治疗组与正常对照组无统计学差异(P>0.05),与高氧对照组具有显著统计学差异(P<0.01)。视网膜铺片正常对照组血管发育成熟,深浅两层血管网结构清晰,可见螺旋动脉,高氧对照组血管迂曲阻塞,大量结构异常新生血管,丧失了血管网结构,GM6001治疗组见两层血管网结构清晰,仍有少部分深层血管阻塞。视网膜免疫组织化学检测GM6001治疗组VEGF及MMP-2与正常对照组有统计学差异(P<0.05),与高氧对照组也有显著统计学差异(P<0.01)。结论:GM6001可抑制视网膜病的血管新生,机制可能是抑制了MMP-2对VEGF的作用。

AIM： To explore the inhibitory effect of GM6001 on retinal neovascularization in neonatal rats with retinopathy. METHODS：Twenty-four 7-day-old Sprague Dawley rats were put into the environment with 750mL/L oxygen for 5 days to establish models of vascular retinopathy. They were subsequently divided into two groups： high oxygen group and treatment group. The treatment group was given GM6001 by injection with the dose of 75μ mol/L 0.5μ L,while the high oxygen control group was not given any intervention. The number of vascular endothelial cell nuclei which extended from retina to vitreous body was counted in the tissue slice of HE staining. The expession of MMP-2 and VEGF was examined by immunohistochemical technology, and the change of reinal blood vessels was investigated by means of retinal preparation on the slide. The normal control group （ 12 rats） was also established. RESULTS： The number of eyes which had neovascularization in GM6001 treated group was more than that in high oxygen group but fewer than that in normal control group. The average number of vascular endothelial cell nuclei which break through the internal limiting membrane on each section was 2.38 ± 0.90, 21.25 ± 1.12 and 1. 23 ± 0.46 respectively in GM6001 treated group, high oxygen group and normal control group. There was no statistical difference between GM6001 treated group and normal control group （ P 〉 0.05）, while there existed statistically significant difference between GM6001 treated group and high oxygen group （ P 〈 0.01 ）, between high oxygen group and normal control group （ P 〈 0. 01 ）. Immunohistochemical examination showed that there was statistical difference between GM6001 treated group and normal control group （ P 〈 0. 05）, and that there was statistically significant difference between GM6001 treated group and high oxygen group （ P 〈 0.01 ） with reagard to the expression of VEGF and MMP-2. In addition, ADPase staining of retina showed that the shallow and deep vasculature was distinctly displayed in normal control group; a lot of curved blocked vessels and retinal neovascularization make the retina lose its original structure in high oxygen group; two layers of vasculature was clearly demonstrated but there was still blockage of deep vessels in GM6001 treated group. CONCLUSION： GM6001 can inhibit retinal neovascularization. The mechanism may involve the interaction between MMP-2 and VEGF.