锌指蛋白基因抑制氧损伤诱导的内皮细胞E-选择素的表达

Zinc finger protein inhibits H_2O_2-inducible E-selectin expression of endothelial cells

目的 :探讨外源性锌指蛋白基因A2 0对氧损伤诱导的内皮细胞E 选择素表达的影响。方法 :DOTAP脂质体介导 pcDNA3 .1EHA2 0质粒转染人脐静脉内皮细胞 ,经G41 8筛选 ,免疫荧光检测A2 0基因的表达 ,原位杂交、免疫组化分别检测过氧化氢诱导的内皮细胞E 选择素表达。结果 :A2 0基因在经G41 8筛选后的内皮细胞中得到高效表达 ,过氧化氢能诱导人内皮细胞E 选择素高表达 ,而A2 0基因能抑制 80 %以上过氧化氢诱导的内皮细胞E 选择素表达 ,两者间相差显著。结论 :A2 0基因能够显著抑制过氧化氢诱导的内皮细胞活化 ,有助于氧损伤的治疗。

Objective:To examine the effectiveness of A20 gene inhibiting H 2O 2-inducible the E-selectin expression of endothelial cells. Methods: With the help of DOTAP, endothelial cells were transfected with pcDNA3.1EHA20. The positive cell clones were selected with G418. The stable transfection and expression of A20 in the endothelial cells were determined by immunofluorescence analysis. The E-selectin expression was checked by immunohistochemical and in situ hybridization analysis. Results:Abundant A20 stable expression in endothelial cells transfected with pcDNA3.1EHA20 were confirmed by immunofluorescence analysis. H 2O 2 significantly increased the expression of E-selectin in endothelial cells. A20 gene inhibited H 2O 2-inducible E-selectin expression by 80%. Conclusion:With the help of DOTAP, hA20 gene can be transferred and stably expressed in endothelial cells, which can inhibit H 2O 2-inducible the E-selectin expression of endothelial cells.