酪氨酸激酶抑制剂（Genistein）在小鼠视网膜新生血管形成中的作用

发布时间：2018-06-12 17:35

本文选题：视网膜新生血管 + 酪氨酸激酶抑制剂　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:研究酪氨酸激酶抑制剂(Genistein)在高氧诱导小鼠视网膜新生血管形成中的作用。方法:应用随机数字表法将36只7日龄清洁级C57BL/6J小鼠随机分为:模型对照组、Gen组、DMSO组、正常组,共四组,每组9只,18眼。前三组小鼠为7日龄小鼠在氧体积分数为(75±2)%的密闭氧箱内生长5天后,返回到正常环境;正常组为自然环境中生长的小鼠。模型对照组和正常组小鼠不给药物处理,Gen组和DMSO组的12天小鼠,玻璃体腔内分别注射Genistein(溶于DMSO,400mg/L)和DMSO各1μl。继续在正常环境下培养小鼠5天后,即小鼠17日龄时,每组两只小鼠4眼行视网膜血管荧光造影铺片观察视网膜血管的形态,各组剩余小鼠全部处死,获取视网膜,应用real-time PCR法,检测小鼠视网膜中VEGF和b FGFm RNA的表达量(2-ΔΔCt);Western blot法,检测各组小鼠视网膜中VEGF和b FGF蛋白的相对表达量(VEGF和b FGF/β-actin)。结果:在模型对照组和DMSO组中,小鼠视网膜中VEGF和b FGFm RNA的表达水平分别为(0.64±0.25和21.40±3.07,0.37±0.23和17.22±2.63),明显高于正常组和Gen组(0.04±0.02和0.67±0.23,0.03±0.02和0.52±0.25),差异有统计学意义(P0.05)。经两两比较,模型对照组和DMSO组、正常组和Gen组比较差异无统计学意义(P0.05),其余各组两两比较,差异均有统计学意义(P0.05)。模型对照组和DMSO组小鼠视网膜中,VEGF和b FGF蛋白的表达水平分别为(1.01±0.05和0.97±0.06,1.06±0.07和1.03±0.08),明显高于正常组和Gen组(0.52±0.05和0.56±0.05,0.73±0.05和0.76±0.07),差异有统计学意义(P0.05)。经两两比较,VEGF,b FGF蛋白在模型对照组和DMSO组中,差异无统计学意义(P0.05),其余两两相比,差异均有统计学意义(P0.05)。视网膜荧光造影铺片显示:模型对照组和DMSO组血管分层和分支较多,血管走行紊乱,新生血管簇明显;Gen组小鼠视网膜血管分层、分支较少,视网膜血管走行相对清晰,未见明显新生血管;正常组视网膜血管走形整齐,清晰,未见新生血管簇。结论:1.高氧诱导小鼠视网膜新生血管中VEGF和b FGF高表达;2.Genistein能抑制视网膜新生血管中VEGF和b FGFm RNA及蛋白的生成;3.Genistein可以抑制视网膜新生血管的形成。
[Abstract]:Aim: to study the role of tyrosine kinase inhibitor Genistein in retinal neovascularization induced by hyperoxia in mice. Methods: Thirty-six C57BL / 6J mice of 7 days old were randomly divided into four groups: model control group (Gen group), normal group (n = 9, n = 18) and control group (n = 9, n = 18). The first three groups were 7-day-old mice, which grew in a closed oxygen box with 75 卤2% oxygen volume fraction for 5 days, then returned to normal environment, while the normal group was the mice growing in the natural environment. The mice in the model control group and the normal group were not treated with drug treatment for 12 days. Genistein (400 mg / L dissolved in DMSOO) and DMSO were injected intravitreally into the vitreous body of mice for 12 days, respectively. After continuing to culture mice in normal environment for 5 days, that is, at the age of 17 days, four eyes of two mice in each group underwent retinal angiography to observe the morphology of retinal vessels, and all the remaining mice in each group were killed to obtain retina. The expression of VEGF and bFGFm mRNA in the retina of mice was detected by real-time polymerase chain reaction. The relative expression of VEGF and bFGF protein in the retina of each group was detected by Western blot method. Results: in the model control group and DMSO group, the expression levels of VEGF and bFGFm mRNA in the retina of mice were 0.64 卤0.25 and 21.40 卤3.07 卤0.37 卤0.23 and 17.22 卤2.63, respectively, which were significantly higher than those in the normal group and Gen group (0.04 卤0.02 and 0.67 卤0.230.03 卤0.02 and 0.52 卤0.25, respectively). There was no significant difference between model control group and DMSO group, normal group and Gen group. The expression levels of VEGF and bFGF in the retina of model control group and DMSO group were 1.01 卤0.05 and 0.97 卤0.06 卤0.07 卤0.07 and 1.03 卤0.08, respectively, which were significantly higher than those in normal group and Gen group, 0.52 卤0.05 and 0.56 卤0.05 卤0.73 卤0.05 and 0.76 卤0.07, respectively. There was no significant difference in VEGFb FGF protein between model control group and DMSO group, but there was significant difference between the other two groups. Fluorescein angiography showed that in the model control group and DMSO group, there were more blood vessel layers and branches, and the blood vessels in the neovascularization group were disordered, the retinal blood vessels in the neovascularization group were obviously stratified, the branches were less, and the retinal blood vessels were relatively clear. There was no obvious neovascularization in the normal group, and no neovascularization was found in the normal group. Conclusion 1. High expression of VEGF and bFGF in retinal neovascularization induced by hyperoxia. 2. Genistein can inhibit the production of VEGF and bFGFm RNA and protein in retinal neovascularization. 3. Genistein can inhibit the formation of retinal neovascularization.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R774.1

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