高氧对小鼠视网膜新生血管模型中根蛋白的影响  

Effects of hyperoxia on radixin in retinal neovascularization model of mice

作　　者：王龙梅[1,2] 闫琳[1,2] 杨侠[2] 董晓光[2] 徐海峰[2] 

机构地区：[1]潍坊医学院,山东省潍坊市261053 [2]山东省眼科研究所青岛眼科医院,山东省青岛市266071

出　　处：《眼科新进展》2014年第3期217-221,225,共6页Recent Advances in Ophthalmology

基　　金：山东省自然科学基金资助(编号:ZR2011HM012)~~

摘　　要：目的探讨高氧诱导C57BL/6J小鼠视网膜新生血管形成过程中根蛋白(Radixin)的表达变化,为进一步研究视网膜新生血管性疾病的防治方法提供理论基础。方法将72只健康7 d龄小鼠随机分为2组,分别为:高氧诱导组:小鼠置于含氧体积分数为75%±2%的氧箱内饲养5 d构建高氧诱导视网膜新生血管动物模型;正常对照组:小鼠置于正常氧环境中饲养。待生后12 d、13 d、17 d、21 d、30 d分别处死小鼠。应异硫氰酸荧光素标记的右旋糖酐(FD-2000S)血管灌注造影视网膜铺片和HE染色观察视网膜新生血管的形态变化;应用免疫组织化学染色检测视网膜中Radixin的表达定位;应用RT-PCR和Westernblot检测不同时间点视网膜Radixin mRNA和蛋白表达情况。结果免疫组织化学染色结果显示两组Radixin均有表达,但高氧诱导组小鼠视网膜新生血管管壁、血管芽、内皮细胞和神经节细胞层Radixin大量表达。Real-time-PCR及Western-blot结果显示,不同组别和不同时间点间Radixin mRNA和蛋白表达水平差异均有统计学意义(mRNA:Fgroup=59.273,P=0.000,Ftime=538.071,P=0.000,Finteraction=297.126,P=0.000;蛋白:Fgroup=419.307,P=0.000,Ftime=663.946,P=0.000,Finteraction=354.538,P=0.000)。高氧诱导组小鼠生后12 d、生后13 d Radixin mRNA和蛋白表达水平均较正常对照组同龄小鼠低,差异均有统计学意义(均为P<0.01),生后17 d、生后21 d Radixin mRNA和蛋白表达水平均较正常对照组同龄小鼠明显增高,差异均有统计学意义(均为P=0.000)。正常对照组Radixin mRNA和蛋白的表达总体稳定在较低水平。两组生后30 d时Radixin mRNA和蛋白表达水平差异均无统计学意义(P=0.058、0.082)。伴随着视网膜新生血管的增生和消退,Radixin mRNA和蛋白的表达水平均呈先升高后降低的趋势。结论 Radixin参与了视网膜新生血管的形成,可能会成为预防和治疗视网膜新生血管性疾病的新靶点。Objective To investigate the expression changes of radixin in oxy- gen-induced retinal（OIR） model of C57BL/5J mice, in order to find new preventive and therapeutical methods for the neovascularization in retinopathy. Methods A total of 72 new born mice with 7 days old were randomly divided into two groups. The mice in OIR group were exposed to hyperoxia for 5 days to establish the OIR model. The oxygen concentration was about 75% ±2%. The mice in normal control group were fed in room air. Mice were put to death at 12 days,13 days,17 days, 21 days and 30 days, respectively. The retinal neovascularization were observed by HE staining and stretched preparation of the retinas after fluorescein isothiocyanate sodium （FD-2000S） intravascular injection. Immunohistochemical staining was used to detect expression location of radixin in retina. Real-time PCR and Western blot were used to detect the expression changes of radixin in retina at gene and protein levels. Results Immunohistochemistry results showed that radixin were expressed in both two group, there were more radixin expressed in retinal neovascularization wall, vascular endotheli- al and ganglion cell layer in the OIR group. RT-PCR and Western blot showed that the expression levels of radixin mRNA and protein were different between groups and dif- ferent time points（ mRNA：Fgroup =59.273 ,P = 0. 000 ,Ftime = 538. 071 ,P = 0. 000, Finteraction = 297. 125 ,P = 0. 000 ; Fgroup = 419. 307, P = 0. 000, Ftime = 553. 945, P = 0. 000, Finteraction = 354. 538 ,P = 0. 000）. The expression levels of radixin mRNA and protein in the OIR group were lower than those in the normal group at 12 days and 13 days（ all P 〈0.01 ） and higher than the normal group at 17 days and 21 days（ all P =0.000）. The expression levels of radixin mRNA and protein in the normal group were very low. There was no difference in radixin mRNA and protein between two groups at 30 days （P = 0. 058, 0. 082 ）. The expression of radixin mRNA and protein were associated with the d

关 键 词：高氧 小鼠 视网膜新生血管 根蛋白 

分 类 号：R587.102[医药卫生—内分泌]