雌二醇对牛视网膜毛细血管内皮细胞VEGF、HIF-1α的调控

山东大学耳鼻喉眼学报

雌二醇对牛视网膜毛细血管内皮细胞VEGF、HIF-1α的调控

马晓华¹，毕宏生¹，李镜海²，解孝锋¹，吴建峰¹，季鹏¹

1. 山东中医药大学眼科中心山东施尔明眼科医院, 济南 250002； 2. 山东大学齐鲁医院眼科，济南 250012

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-02-24 发布日期:2008-02-24
通讯作者: 马晓华

Effect of 17β-estradiol on VEGF and HIF-1α mRNA expressions in bovine retinal vascular endothelial cells

MA Xiao-hua¹，BI Hong-sheng¹， LI Jing-hai²， XIE Xiao-feng¹，　WU Jian-feng¹，JI Peng¹

1. Eye Center of Shandong University of Traditional Chinese Medinice，Shandong Shierming Eye Hospital, Jinan 250002;2. Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan 250012, China

Received:1900-01-01 Revised:1900-01-01 Online:2008-02-24 Published:2008-02-24
Contact: MA Xiao-hua

摘要/Abstract

摘要： 目的观察在不同氧环境下，雌二醇（17β-estradiol，E2）对培养的牛视网膜毛细血管内皮细胞(bovine retinal vascular endothelial cells， BRECs) 血管内皮生长因子（vascular endothelial growth factor ，VEGF）mRNA、低氧诱导因子-1（hypoxia-inducible factor-1α，HIF-1α）mRNA的影响。方法以培养的BRECs为模型，在正常氧及低氧（1% O2）条件下，给予不同生理浓度的E2及雌激素受体拮抗剂他莫昔芬，作用不同时间（8、24h），RT-PCR检测VEGF、HIF-1α mRNA的表达。结果① 低氧条件下，BRECs的HIF-1α、VEGFmRNA表达较正常氧条件下明显增多（P＜0.05），且VEGF、HIF-1α mRNA两者之间有明显的正相关性（r=0.82，P＜0.05）；②　正常氧条件下，给予E2（10-12,10-10,10-8mol/L）作用24h，E2呈浓度依赖性促进BRECs的VEGF mRNA表达（P＜0.05)。10-8mol/L E2作用8、24h，VEGFmRNA表达量呈时间依赖性增多（P＜0.05)；而E2不影响HIF-1α mRNA的含量（P＞0.05）；③　低氧条件下，给予E2（10-12,10-10,10-8mol/L）作用24h，E2呈浓度依赖性抑制BRECs的HIF-1α、VEGF mRNA表达（P＜0.05）。10-8mol/L E2作用8、24h，HIF-1α、VEGF mRNA表达量呈时间依赖性降低（P＜0.05）；④ 他莫昔芬可逆转E2的作用。结论生理浓度的E2对VEGF mRNA具有双重调控作用：正常氧条件下促进BRECs VEGF表达；低氧条件下通过HIF-1α降低VEGF表达，并呈剂量、时间依赖性。

关键词: 雌二醇, 视网膜血管, 内皮细胞, 血管内皮生长因子, 低氧诱导因子-1α

Abstract: Objective To investigate the effect of the physiological level of estradiol on mRNA expressions of the vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) in bovine retinal vascular endothelial cells (BRECs) under different oxygen conditions. MethodsmRNA expressions of VEGF and HIF-1α were determined by reverse transcription polymerase chain reaction (RTPCR) in BRECs, with E2（10-12,10-10,10-8mol/L）or phosphatebuffered saline (PBS) and estrogen receptor antagonist tamoxifen (10-7mol/L) under normoxia and hypoxia after 8 hours and 24 hours. Results① Under hypoxia, gene expressions of VEGF and HIF-1α mRNA in BRECs more significantly increased than in the controls(P＜0.05). There were evident positive correlations between the expressions of VEGF mRNA and HIF-1α mRNA（r=0.82）. ② After BRECs were treated with the indicated concentrations of E2 under normoxia for 24 hours, the expression of VEGF mRNA increased in a dose-dependent manner between 10-12～-8mol/L (P＜0.05). After BRECs being treated with 10-8mol/L E2 8 hours and 24 hours, the expression of VEGF mRNA was increased in a time-dependent manner(P＜0.05). Whereas E2 did not affect the expression of HIF-1αmRNA（P＞0.05）. ③ After BRECs were treated with the indicated concentrations of E2 under hypoxia for 24 hours, the expressions of HIF-1α mRNA and VEGF mRNA decreased in a dose-dependent manner between 10-12～-8mol/L(P＜0.05). After BRECs were treated with　10-8mol/L E2 for 8 hours and 24 hours, the expressions of HIF-1α mRNA and VEGF mRNA decreased in a timedependent manner (P＜0.05). ④ An overdose of tamoxifen (10-7mol/L) reversed the effect of E2(10-8mol/L) (P＜0.05). ConclusionsThe physiological level of E2 has a contrast effect on VEGF mRNA in a dose and timedependent manner based on oxygen conditions. E2 increases the gene expression of VEGF in BRECs under normoxia, whereas E2 decreases the gene expression of VEGF in BRECs through HIF-1α under hypoxic conditions.

Key words: Retinal vessels, Endothelial cells, 17β-estradiol, Vascular endothelial growth factor, Hypoxia-inducible factor-1α

中图分类号:

R774

马晓华,毕宏生,李镜海,解孝锋,吴建峰,季鹏 . 雌二醇对牛视网膜毛细血管内皮细胞VEGF、HIF-1α的调控[J]. 山东大学耳鼻喉眼学报, 2008, 22(1): 6-09 .

MA Xiao-hua,BI Hong-sheng,LI Jing-hai,XIE Xiao-feng,WU Jian-feng,JI Peng . Effect of 17β-estradiol on VEGF and HIF-1α mRNA expressions in bovine retinal vascular endothelial cells[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2008, 22(1): 6-09 .

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