摘要
目的探讨在缺氧状态下视网膜血管内皮细胞缺氧诱导因子-1α(HIF-1α)及其mRNA表达的时空规律。方法对分离的牛视网膜血管内皮细胞分别进行常规和CoCl2模拟缺氧培养,采用免疫组织化学和逆转录多聚酶链反应(RT-PCR)检测不同缺氧时间HIF-1α及其mRNA的表达。结果正常对照组HIF-1α有极低表达,缺氧1h表达量显著上升,4h达到高峰,16h后下降。与对照组比较,各缺氧组HIF-1α的表达均有显著统计学差异(P<0·01)。但各缺氧组HIF-1αmRNA的表达无显著统计学差异。结论视网膜血管内皮细胞在缺氧早期HIF-1α表达有短暂、迅速的增加,但其mRNA的表达无明显变化,提示缺氧可以促进视网膜血管内皮细胞HIF-1α的表达,但不是在基因的转录水平。
Objective Vascular endothelial growth factor（VEGF） is thought to be a critical factor to induce new blood formation. The study on hypoxia inducible factor（ HIF-1 α）in human＇s other organs showed that HIF-1 α is a modulating and controlling factor of vascular endothelial growth factor（VEGF） , however, the effect of HIF-1 α on retinal vascular endothelial cells in low oxygen status is still unknown. The purpose of this study was to investigate the sequential changes of HIF-1 α protein and mRNA in hypoxic bovine retinal vascular endothelial cells. Methods The bovine retinal vascular endothelial cells were cultured in normoxic condition （normal control group） or 125 μmmol/L CoCl2-induced hypoxic condition （hypoxic group） respectively. Expressions of HIF-1 α protein were measured with immunohistochemical staining, and RT-PCR was used to determine expressions of the HIF-1 α mRNA at 1,2,4,8,16 hours after culture. Results HIF-1 α began to increase 1 hour after hypoxia（56. 18 ± 2.76） and reached the peak at 4 hours（ 134.08 ± 2.94） and declined significantly 16 hours（99. 50 ± 2.11）later. Compared with the normoxic group （ 30. 94 ± 1.72） , the expressions of HIF-1 α in the all hypoxic groups had significant difference（P 〈 0. 01 ）. RT-PCR revealed that HIF-1 α mRNA was expressed in the hypoxie group and normoxie group. There was not significant difference between the hypoxic group and normoxic group （ P 〉 0.05 ）. Conclusion HIF-I ot is overexpressed in retinal vascular endothelial cells in hypoxic status,otherwise,hypoxia induce time- dependent changes of HIF-1 α protein expression,but this alteration is not modulated in the transcription level.
出处
《眼科研究》
CSCD
北大核心
2006年第4期356-358,共3页
Chinese Ophthalmic Research
