高氧对小鼠视网膜新生血管模型中根蛋白的影响

发布时间：2018-11-02 08:45

【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P0.01),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。
[Abstract]:Objective to investigate the changes of root protein (Radixin) expression during retinal neovascularization in C57BL/6J mice induced by hyperoxia, and to provide a theoretical basis for further study on the prevention and treatment of retinal neovascularization diseases. Methods Seventy-two healthy 7-day-old mice were randomly divided into two groups: hyperoxia-induced group: mice were fed in oxygen box containing 75% 卤2% oxygen for 5 days to establish hyperoxia-induced retinal neovascularization animal model; Normal control group: mice were fed in normal oxygen environment. The mice were killed on the 12th day, 13th day, 17 day, 21 day and 30 d after birth. The morphological changes of retinal neovascularization were observed by fluorescein isothiocyanate labeled dextran (FD-2000S) angiography and HE staining, and the expression of Radixin in the retina was detected by immunohistochemical staining. RT-PCR and Westernblot were used to detect the expression of Radixin mRNA and protein in retina at different time points. Results Immunohistochemical staining showed that Radixin was expressed in both groups, but Radixin was expressed in retinal neovascularization wall, vascular bud, endothelial cells and ganglion cell layer in hyperoxia-induced group. Real-time-PCR and Western-blot showed that the expression of Radixin in retinal neovascularization vessel wall, vascular bud, endothelial cell and ganglion cell layer was higher than that in control group. There were significant differences in the expression of Radixin mRNA and protein between different groups and different time points (mRNA:Fgroup=59.273,P=0.000,Ftime=538.071,P=0.000,Finteraction=297.126,P=0.000;). Protein: Fgroup=419.307,P=0.000,Ftime=663.946,P=0.000,Finteraction=354.538,P=0.000). The expression levels of d Radixin mRNA and protein in hyperoxia-induced group were significantly lower than those in normal control group at 12 days after birth and 13 days after birth (all P0.01), and 17 days after birth, the expression levels of protein and protein in hyperoxia-induced group were significantly lower than those in normal control group (P0.01). The expression levels of 21 d Radixin mRNA and protein in postnatal mice were significantly higher than those in the control group (P 0. 000). The expression of Radixin mRNA and protein was stable in normal control group. There was no significant difference in the expression of Radixin mRNA and protein between the two groups 30 days after birth (P0. 058 and 0. 082). With the proliferation and regression of retinal neovascularization, the expression of, Radixin mRNA and protein increased first and then decreased. Conclusion Radixin is involved in the formation of retinal neovascularization and may be a new target for the prevention and treatment of retinal neovascular diseases.
【作者单位】：潍坊医学院;山东省眼科研究所青岛眼科医院;
【基金】：山东省自然科学基金资助(编号:ZR2011HM012)~~
【分类号】：R774.1

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