摘要
目的探讨基质金属蛋白酶抑制剂-1(TIMP-1)对糖尿病小鼠视网膜的保护作用及机制。方法将96只6~8周龄C57bl小鼠随机分为对照组、糖尿病组和TIMP-1干预组[糖尿病造模后第1和2个月玻璃体腔注射5μL TIMP-1(1 mg/L)各1次]。5个月后进行视网膜灌注铺片检测渗漏,Western blot和RT-q PCR分别检测基质金属蛋白酶9(MMP-9)、血管内皮生长因子(VEGF)、肿瘤坏死因子α(TNF-α)和环氧合酶-2(COX-2)mRNA及蛋白表达,用生化法检测视网膜组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量。结果对照组视网膜无渗漏,糖尿病组视网膜渗漏明显高于TIMP-1干预组(P〈0.01)。糖尿病组视网膜MMP-9、VEGF、TNF-α、COX-2 mRNA与蛋白表达水平以及MDA含量均明显高于对照组(P〈0.01),TIMP-1干预组则显著回降(P〈0.01),但仍高于对照组(P〈0.01);糖尿病组视网膜SOD、GSH-Px活性明显低于对照组(P〈0.01),TIMP-1干预组则显著回升(P〈0.01),但仍低于对照组(P〈0.01)。结论 TIMP-1能够通过减少氧化应激与VEGF表达对糖尿病小鼠视网膜起保护作用。
Objective To investigate the protective effect and mechanism of matrix metalloproteinase inhibitor-1( TIMP-1) against diabetic retinopathy in mice. Methods A total of 96 old C57 bl mice of 6- 8 weeks were randomly divided into 3 groups: control group,diabetic group and TIMP-1 group( mice were intravitreously injected with TIMP-1 5 μL( 1 μg/m L) in the first and second months respectively after diabetes-induced). Retinal vascular leakages were investigated by retinal perfusion of evans blue after 5 months. The mRNA and protein expression levels of metalloproteinase 9( MMP-9),vascular endothelial growth factor( VEGF),tumor necrosis factor alpha( TNF-α) and epoxy synthase-2( COX-2) were also analyzed by RT-q PCR and Western blot. The superoxide dismutase( SOD),glutathione peroxidase( GSH-Px) activity and malondialdehyde( MDA) content in the retina tissue were also detected with bio-chemical method. Results There was no retinal leakage in control group,but diabetic group was significantly higher than TIMP-1 group( P〈 0. 01). The mRNA and protein expression of MMP-9,VEGF,TNF-α,COX-2 and the MDA content in diabetic group were significantly higher than control group( P〈 0. 01),while decreased in TIMP-1 group( P〈 0. 01) and also higher than control group( P〈 0. 01). The SOD and GSH-Px activity in diabetic group were significantly lower than control group( P〈 0. 01),while in TIMP-1group they increased( P〈 0. 01) and also lower than that of control group( P〈 0. 01). Conclusions TIMP-1 can protect the retina of diabetic mice by reducing the oxidative stress and the expression of VEGF.
出处
《基础医学与临床》
CSCD
2016年第12期1687-1692,共6页
Basic and Clinical Medicine
基金
三峡大学青年科学基金(KJ2012A019)