高氧诱导小鼠视网膜新生血管过程中不同细胞因子mRNA表达水平的检测

发布时间：2018-03-18 12:15

本文选题：视网膜新生血管　切入点：血管内皮生长因子　出处：《浙江大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的研究高氧诱导小鼠视网膜新生血管发生过程中血管内皮生长因子(VEGF),成纤维细胞生长因子(bFGF),内皮抑素(ET)和乙酰肝素酶(HPA)在视网膜中的表达。方法实验组将21只新生7天幼鼠放于体积分数约75%的高氧环境中饲养5天,再置于正常环境中饲养。另外21只新生幼鼠在正常环境中饲养作为正常对照组。分别在小鼠出生后第12、17、25天通过高分子量荧光素血管造影观察视网膜血管形态变化。并取小鼠视网膜,通过逆转录聚合酶链反应(RT-PCR)分别检测不同时间点视网膜组织中VEGF, bFGF, ET, HPA mRNA水平的变化,比较高氧组与对照组的差异。结果高氧组小鼠在生后12天时主要表现为后级部大片无灌注区,视网膜血管管径变细,分支减少,仅在周边部网膜形成少量血管网,正常对照组血管分布均匀,走形正常；而在生后17天时,高氧组小鼠血管走形迂曲紊乱,可见大量新生血管存在,对照组血管则均匀分布于整个视网膜；生后25天时高氧组血管分布较前均匀,但仍存在一定量的新生血管。另外生后3个不同时间点高氧组VEGF、bFGF和HPA mRNA表达水平逐渐增高,与对照组比较差异有统计学意义(F=113.608,P=0.000,F=59.235,P=0.OOO,F=188.493,P=0.000)。而ETmRNA表达水平逐渐降低,与对照组比较差异有统计学意义(F=32.365,P=0.000)。结论应用75%浓度的高氧可成功诱导小鼠视网膜新生血管形成,在新生血管形成过程中VEGF、bFGF、HPA作为进视网膜新生血管生长的重要因子,其mRNA表达水平升高；而ET作为抑制视网膜新生血管生长的细胞因子,其mRNA表达水平降低。
[Abstract]:Purpose. To study the expression of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), endostatin (et) and heparanase (HPA) in the retinal neovascularization of mice induced by hyperoxia. Method. In the experimental group, 21 newborn 7-day-old rats were fed in a hyperoxic environment with a volume fraction of about 75% for 5 days. The other 21 newborn rats were fed in normal environment as normal control group. The morphological changes of retinal blood vessels were observed by high molecular weight fluorescein angiography on the 25th day after birth. And the retina of mice, The changes of VEGF, bFGF, et, HPA mRNA in retinal tissues at different time points were detected by reverse transcriptase polymerase chain reaction (RT-PCR), and the difference between hyperoxia group and control group was compared. Results. At 12 days after birth, the hyperoxia group showed a large area of no perfusion in the posterior part, the diameter of retinal vessels became thinner, the branches decreased, only a small number of vascular networks were formed in the peripheral omentum, while in the normal control group, the vascular distribution was uniform and the shape was normal. On the 17th day after birth, the hyperoxia group was found to have a large number of neovascularization, while the control group was evenly distributed throughout the retina, and on the 25th day after birth, the vascular distribution in the hyperoxia group was more uniform than that in the former. However, there was still a certain amount of neovascularization. The expression of VEGF bFGF and HPA mRNA in hyperoxia group gradually increased at three different time points after birth, and the difference was statistically significant compared with the control group. The expression level of ETmRNA decreased gradually, and the expression level of VEGFN bFGF and HPA mRNA in hyperoxia group was lower than that in the control group (P < 0.05), but the expression level of VEGFO bFGF and HPA mRNA in hyperoxia group was gradually decreased, compared with that in control group (P < 0.05). Compared with the control group, the difference was statistically significant. Conclusion. Hyperoxia at the concentration of 75% could successfully induce retinal neovascularization in mice. During the process of neovascularization, the expression level of mRNA was increased as an important factor for the growth of retinal neovascularization. Et, as a cytokine to inhibit retinal neovascularization, decreased the expression of mRNA.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.1

【参考文献】