基质金属蛋白酶抑制剂GM6001对糖尿病大鼠血-视网膜屏障的影响  

Effect of matrix metalloproteinase inhibitor GM6001 on blood-retina barrier permeability in diabetic rats

作　　者：顾永昊[1] 柯根杰[1] 王林[1] 何勤[1] 周恩亮[1] 

机构地区：[1]安徽医科大学附属省立医院眼科,合肥230001

出　　处：《中华实验眼科杂志》2014年第11期1010-1013,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：安徽省自然科学基金项目（1208085QH167）

摘　　要：背景 研究表明,多种基质金属蛋白酶（MMPs）家族成员在糖尿病视网膜病发（DR）的发病过程中发挥作用,但是否MMPs抑制剂能改善MMPs对血-视网膜屏障（BRB）的破坏作用尚不十分清楚.目的 探讨人工合成MMPs抑制剂GM6001对糖尿病大鼠BRB的影响.方法 24只成年清洁级SD大鼠按随机数字表法随机分为对照组、糖尿病组和糖尿病＋GM6001组.采用链脲佐菌素诱导腹腔内注射法诱导大鼠糖尿病模型,对照组以相同的方法注射等体积枸橼酸盐缓冲液.造模成功后3d和14d,糖尿病＋GM6001组大鼠玻璃体腔内注射10μl（100tμmol/L）GM6001各1次,对照组和糖尿病组大鼠以相同的方法注射等体积生理盐水.注射后1个月摘取大鼠一侧眼球,采用逆转录PCR（RT-PCR）法检测大鼠视网膜中MMP-2 mRNA和MMP-9 mRNA的相对表达量;用伊文思蓝（EB）灌注大鼠右侧颈静脉,120 min后以约120 mmHg（1 mmHg=0.133 kPa）的灌注压于大鼠左心室灌注枸橼酸钠缓冲液配制的质量分数1％多聚甲醛溶液,2 min后摘取大鼠另一侧眼球,观察大鼠视网膜EB的渗漏量.结果 RT-PCR检测显示MMP-2、MMP-9和GAPDH的反应条带分别位于436、536和484 bp.3个组大鼠视网膜中MMP-2 mRNA和MMP-9 mRNA相对表达量总体差异均有统计学意义（F=20.336,P=0.000;F=8.742,P=0.002）;其中糖尿病组和糖尿病＋GM6001组大鼠视网膜中MMP-2 mRNA和MMP-9 mRNA的相对表达量明显高于对照组大鼠,差异均有统计学意义（P＜0.01）,而糖尿病＋GM6001组大鼠视网膜中MMP-2 mRNA和MMP-9 mRNA的相对表达量明显低于糖尿病组大鼠,差异均有统计学意义（P=0.01、0.02）.对照组、糖尿病组和糖尿病＋GM6001组大鼠视网膜中标准化EB质量分数分别为（12.60±3.50）、（26.52±7.14）和（17.55±2.65） ng/mg,总体差异有统计学意义（F=17.032,P＜0.01）,其中糖尿病组大鼠视网膜中标准化EB质量分数值明显高于对照组,而糖尿病＋GM6001组较糖�Background Studies showed that some members of matrix metalloproteinases （MMPs） play an important role in the pathogenesis of diabetic retinopathy （DR）.However,whether MMPs inhibitor can deter blood retinal barrier （BRB） from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes＋GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes ＋ GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 （100 μ mol/L） was intravitreously injected in the third and fourteenth day after modeling in the diabetes＋ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR （RT-PCR）.Evens blue （EB） was infused via the right jugular vein,and paraformaldehyde solution 1% was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant （F =20.336,P =0.000 ; F =8.742,P =0.002） ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes ＋GM6001 group than those in the control group （all at P〈0.01）,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes＋GM6001 group was signific

关 键 词：金属内肽酶类/拮抗剂和抑制剂 糖尿病/并发症 视网膜 血-视网膜屏障 渗透性 动物模型 SD大鼠 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]