【摘要】 目的探讨缺氧诱导因子-1α(HIF-1α)反义寡核苷酸(ASODN)治疗视网膜新生血管性疾病的可行性及机制。方法将体外培养的牛眼视网膜微血管内皮细胞(BREC)分为转染组和未转染组,转染组体外转染HIF-1α ASODN(根据GenBank提供的HIF-1αcDNA序列设计),两组均予125μmol/L的CoCl2行缺氧处理。采用免疫荧光法检测细胞HIF-1α的表达;ELISA法检测细胞培养上清液中VEGF蛋白水平。结果脂质体介导的HIF-1α ASODN可成功转染BREC细胞。未转染组缺氧开始时HIF-1α有极低表达,缺氧1h时表达量显著升高,4h达到高峰,16h后下降。而转染组HIF-1α的表达始终在极低水平,两组间有显著性差异(P<0.01)。转染组从缺氧1h开始VEGF表达明显低于未转染组(P<0.01)。结论脂质体介导的HIF-1α ASODN能有效抑制BREC细胞HIF-1α的表达,从而降低细胞VEGF的表达。更多还原

【Abstract】 Objective To investigate the feasibility and mechanism of hypoxia-inducible factors-1α( HIF-1α) antisense oligodeoxynucleotides( ASODN) for the therapy of retinal neovasculation. Methods Bovine hypoxic retinal endothelial cell ( BREC) were cultured in vitro and divided into transfection group and non-transfection group,the transfection group was transfected with HIF-1α ASODN,both the two groups were given CoCl2-induced hypoxic treatment. Expression of HIF-1α was measured with immuno fluorescence staining,VEGF of BREC cells was detected by ELISA assay. Results HIF-1α ASODN was successfully transfected into BREC cells. The expression of HIF-1α in non-transfection group was very low at the early time of anoxia,increased at 1h ,reached peak at 4h ,decreased after 16h. The expression of HIF-1αin transfection group maintained at very low level and showed significant difference to the nontransfection group( P < 0. 01) . Conclu- sions The transfection of HIF-1α ASODN can effectively down-regulate the expression of HIF-1α and VEGF. The therapeutic prospect of HIF-1α ASODN on retinal neovasculation is worthy of further investigation.更多还原