熊果酸对小鼠视网膜缺血模型中 VEGF、COX-2及 MMP-2表达的影响  

The influence of ursolic acid on VEGF, COX-2 and MMP-2 expressed in the mouse retinal ischemic model

作　　者：贺玲[1] 郑晓龙[1] 王晓[2] 温中华[1] 郭晶晶[1] 

机构地区：[1]解放军第三七一中心医院眼科,河南省新乡453000 [2]新乡医学院

出　　处：《中国医师杂志》2014年第4期443-446,共4页Journal of Chinese Physician

基　　金：河南省新乡市重点科技攻关计划基金项目（ZG12013）

摘　　要：目的：观察熊果酸对氧诱导小鼠视网膜组织中血管内皮细胞生长因子（VEGF）、环氧化酶-2（COX-2）及基质金属蛋白酶-2（MMP-2）表达的影响，从而探讨熊果酸（UA）的抗新生血管机制。方法将60只7d龄清洁级C57BL/6J小鼠，按随机数字表法分为6组：模型对照组（PBS液）、阳性对照组（曲安奈德）、UA干预组（低、中、高剂量）和空白对照组，每组10只小鼠（各20只眼）。空白对照组小鼠在空气中喂养，其它各组制备缺氧小鼠视网膜新生血管模型。模型成功后，各组立即给予相应的药物治疗：模型对照组小鼠玻璃体腔注射无菌PBS液3μl，UA干预组（低、中、高剂量）小鼠分别于玻璃体腔注射1.5、3.0、6.0μgUA各3μl，阳性对照组小鼠玻璃体腔注射曲安奈德（1ml∶40mg）3μl。17d龄时过量麻醉处死各组小鼠，摘除眼球，制作视网膜组织病理切片，利用免疫组化方法检测视网膜组织中VEGF、COX-2及MMP-2的阳性表达；制备视网膜组织匀浆液，应用Westernblot方法分别检测每组视网膜组织中VEGF、COX-2及MMP-2的阳性表达和蛋白测定；以及应用RT-PCR法分别检测每组视网膜组织中VEGF、COX-2及MMP-2的mRNA表达。结果不同组别视网膜组织中COX-2、VEGF和MMP-2阳性表达、蛋白测定和mRNA的表达比较，模型对照组与空白对照组比较，各细胞因子均明显升高（P＜0.05）；高剂量UA干预组与模型对照组比较明显降低（P＜0.05）；高剂量UA干预组与阳性对照组比较差异无统计学意义（P＞0.05）。高剂量UA干预组VEGF、COX-2及MMP-2的表达明显少于低剂量UA干预组（P＜0.05）。结论UA抑制小鼠视网膜缺血模型中VEGF、COX-2及MMP-2阳性表达、蛋白测定和mRNA的表达，从而对视网膜新生血管的形成有抑制作用，并与UA的剂量呈正相关。Objective To investigate the influence of ursolic acid on vascular endothelial growth factor （ VEGF） , cycloxygen-ase-2 （COX-2）, and matrix metalloproteinases-2 （MMP-2） expressed in the mouse retinal ischemic model , and to explore the mecha-nisms of anti-angiogenesis.Methods Sixty 7-day clean-class C57BL/6J mice were divided randomly into 6 groups [ n =10 mice （20 eyes） per group]：blank control, model control （PBS）, positive control （triamcinolone）, and ursolic acid （UA） intervention （low-dose, medium-dose, and high-dose）.Mice in the blank control group were raised in air , and mice in other groups in（75%＆#177;2%）O2 high-oxygen environment for 5 consecutive days .Mice in the model control group and breastfeeding mice were put back in air environ-ment （21%O2 ） on the 12th day after the new-born mice to induce the generation of retinal neovascularization .When models were suc-cessful, the drug treatments were applied immediately to the corresponding groups , with injection of 3μl of sterile PBS in model control group, 3 μl of 1.5, 3.00 and 6.0 μg UA in UA intervention group, and 3 μl of triamcinolone （1 ml∶40 mg） in positive control group, respectively.All mice were killed after overdose anesthesia on the 17th day.Their eyeballs were made into samples and retinal tissue pathological sections with H-E dying method.The positive expressions of VEGF , COX-2, and MMP-2 were detected with immu-nohistochemical method .The fresh retinal tissue homogenate was prepared to detect the protein expressions of VEGF , COX-2, and MMP-2 in retinal tissue with western blot method ,and mRNA expressions of VEGF , COX-2, and MMP-2 were detected with real-time fluorescent quantitative polymerase chain reaction （ RT-PCR） .Results According to protein and mRNA expressions of VEGF , COX-2,and MMP-2 in retinal tissue among six groups , protein expressions of VEGF , COX-2, and MMP-2 in model group were significantly＆amp;nbsp;higher than those in blank group （ P ＆lt;0.05 ） .Eac

关 键 词：熊果酸 药理学 缺血 视网膜疾病 血管内皮生长因子类 代谢 基质金属蛋白酶2 代谢 前列腺素内过氧化物合酶类 生物合成 

分 类 号：R965[医药卫生—药理学]