摘要
目的探讨视网膜新生血管发生过程中Twist基因的表达情况。方法对照实验研究。将40只新生C57／BL小鼠分为两组：（1）对照组，正常空气环境中饲养；（2）高氧组，在高氧环境中制作小鼠缺氧性视网膜新生血管动物模型。将出生第7天的小鼠放入氧箱，在75％浓度的高氧环境中饲养5d后，取出氧箱。待出氧箱第12天和第17天（视网膜新生血管发生高峰时间）分别处死小鼠，制作视网膜铺片和切片，观察小鼠视网膜新生血管发生情况。应用免疫组织化学染色检测小鼠视网膜血管内皮细胞（VE）-钙黏蛋白、波形蛋白及Twist基因表达变化情况；提取全视网膜RNA，应用逆转录聚合酶链反应（RT-PCR）法检测VE-钙黏蛋白、波形蛋白及Twist基因表达变化情况。应用SPSS11．5统计学软件进行数据处理，对高氧组与对照组小鼠出生后不同时间的VE．钙黏蛋白、波形蛋白、Twist基因表达的灰度值比较，采用两因素析因设计的方差分析，以P〈0．05作为差异有统计学意义。结果免疫组织化学检测结果显示，出生后第17天的小鼠，高氧组VE一钙黏蛋白表达的灰度值（65．19±8．39）与对照组（75．36±7．04）相比明显减少（F=8．616，P=0．009），波形蛋白表达的灰度值（95．09±14．13）与对照组（82．14±6．32）相比明显升高（F=6．999，P=0．016），Twist基因表达的灰度值（119．48±7．90）与对照组（93．30±6．37）相比亦明显升高（F=66．557，P=0．000）。RT-PCR检测结果显示，不同处理组间波形蛋白、Twist基因表达的灰度值与检查时间存在交互作用（F=5．508，P=0．032；F=17．760，P=0．001）。结论在小鼠视网膜新生血管形成过程中，细胞转型调控的Twist基因发挥着重要作用，其作用机制可能是通过下调血管内皮细胞间的紧密连接蛋白，促进内皮细胞转型实现。
Objective To study the mechanisms of the Twist gene and cell transition during the angiogenesis in a mouse model of oxygen induced retinopathy. Methods It was a experimental study. 40 new born C57/BL mice were divided into two groups ; the control group, which were fed in the room air, and the study group, which were fed 7 days in the normal environment, followed by exposure to hyperoxia （75% O2） for 5 days. The mice were sac,fficed at post born 12 day and post born 17 day. The Twist and VE-cadherin expression were detected with immunohistochemistry. The total retinal RNA was extracted and the expression level of the VE-cadherin, vimentin and twist were examined with RT-PCR method. Results Immunohistochemistry proved there was no significant difference at post born 12 day. At post born 17 day, compared with the control group（75.36 ± 7.04） ,VE-cadherin expression of hyperoxia group（65.19 ± 8.39） decreased （ F = 8. 616, P = 0. 009）. Compared with the control group （ 82.14 ± 6. 32 ）, Vimentin expression of hyperoxia group （ 95.09 ± 14. 13 ） increased, compared with the control group （ 93.30 ± 6. 37 ）, Twist expression of hyperoxia group （ 119. 48 ± 7.90 ） increased （ F = 66. 557, P = 0. 000 ） significantly. RT-PCR examination revealed there exist the interaction between Vimentin, Twist expression and the detection time （ F = 5. 508, P = 0. 032 ; F = 17. 760, P = 0. 001 ; respectively）. At post born 12 day, compared with the control group （ 0. 77 ± 0. 10 ）, VE-cadherin expression of hyperoxia group （ 0. 64 ± 0.09 ） decreased （ P =0. 047 ）. Compared with the control group （ 0. 24 ± 0. 05 ）, Vimentin expression of hyperoxia group （ 0. 39 ± 0. 09）increased（P =0. 033）. At post born 17 day, compared with the control group（0. 75 ±0. 12）, VE- cadherin expression of hyperoxia group （ 0. 51 ± 0. 07 ） decreased more obviously （ P = 0. 002 ）, compared with the control group（ 0. 36 ± 0. 06） , Vimentin expression of hyperoxia group（ 0. 70 ± 0. 14）increased （P = 0. 000）. Compared with the control group （ 0. 89 ± 0. 11 ）, Twist expression of hyperoxia group （ I. 24 ± 0. 15 ） increased（ P = 0. 003 ）. Conclusion Twist, as a cell transition gene, participated in the angiogenesis of the oxygen induced retinopathy, and that the Twist induced endothelium-mesenchymal transition may be one of the main reasons.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2008年第7期634-639,共6页
Chinese Journal of Ophthalmology
基金
国家自然科学基金资助项目（30772386）
山东省自主创新重大科技专项计划基金资助项目（2006GG1102020）
山东省自然科学基金资助项目（Y2006C94）
关键词
视网膜新生血管化
TWIST转录因子
内皮细胞
基因表达调控
细胞分化
Retinal neovascularization
Twist transcription factor
Endothelial cells
Gene expression regulation
Cell differentiation
作者简介
（刘廷为在读博士生）
通讯作者：董晓光，Email：lixinxie@public．qd．sd．cn
