HIF-1α反义寡核苷酸脂质体的制备和对牛视网膜血管内皮细胞HIF-1α表达的影响

Preparation of Liposomes/HIF-1α ASODN Complexes and Their Inhibiting Effect on HIF-1α Expression in Bovine Retinal Endothelial Cell

【目的】应用 HIF- 1α反义寡核苷酸(ASODN)处理牛眼视网膜微血管内皮细胞,观察细胞摄取ASODN 的情况和对缺氧时细胞HIF 1α表达的影响。【方法】对分离的牛视网膜血管内皮细胞进行 HIF -1α反义寡核苷酸的转染, CoCl2 模拟缺氧培养,采用免疫组化检测不同缺氧时间 HIF 1α的表达。【结果】未转染组在缺氧1 h时HIF 1α表达量显著上升,4 h达到高峰,16 h后下降。ASODN转染组在各时相HIF 1α表达量均较低,两组间HIF- 1α的表达有显著性差异(P<0.01)。【结论】以脂质体为载体能高效率地将HIF- 1αASODN转染至视网膜血管内皮细胞内,明显抑制 HIF- 1α蛋白质的表达,这可能为 HIF- 1αASODN用于视网膜新生血管的治疗提供理论依据。

To study inhibition of HIF-1α expression in bovine retinal endothelial cell(BREC) by HIF-1α fully phosporothioated antisense oligodeoxynucleotide (HIF-1α ASODN).BRECs were incubated with HIF-1α ASODN and were cultured in COCl 2-induced hypoxic condition.HIF-1α expression was detected with immunohistochemistry.HIF -1α began to increase 1h after hypoxia, and reached the peak at 4h. 16h later , it declined significantly. HIF-1α antisense oligonucleotides obviously inhibited HIF-1α expression in BREC.[Conclusion]HIF-1α antisense oligonucleotides can suppress HIF-1α expression of bovine retinal endothelial cell effectively. It has good prospect in resisting retinal angiogenesis treatment.