转染锌指蛋白基因A20抑制内毒素诱导的内皮细胞IL-8表达的研究

目的探讨转染外源性锌指蛋白基因A20对内毒素诱导的内皮细胞白细胞介素-8(IL-8)表达的影响.方法DOTAP脂质体介导pcDNA3.1EHA20质粒转染人脐静脉内皮细胞,经G418筛选,免疫荧光检测A20基因的表达,双抗夹心EIISA法检测内皮细胞分泌IL-8的含量变化.结果A20基因在经G418筛选后的内皮细胞中得到高效表达;内毒素能刺激人内皮细胞分泌IL-8;A20基因能减少70%以上内毒素诱导的内皮细胞IL-8的分泌,两者间相差显著(P＜0.05).结论转染外源性A20基因能够显著抑制内毒素诱导的内皮细胞活化,有助于炎症的治疗.

Zinc finger protein gene A20 inhibits lipopolysaccharide-induced IL-8 expression of endothelial cells

Objective To investigate the effects of zinc finger protein gene A20 on the inhibition of lipopolysaccharide (LPS) induced interleukin 8 (IL 8) expression in endothelial cells. Methods Plasmid pcDNA3.1EHA20 was transfected into human umbilical vein endothelial cells (HUVECs) by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of A20 in HUVECs were determined by immunofluorescent analysis. IL 8 expression was detected by sandwich ELISA with double monoclonal antibodies. Results High expression of A20 gene in HUVECs transfected with pCDNA3.1EHA20 was confirmed by immunofluorescent analysis. IL 8 expression increased in LPS induced endothelial cells. A20 gene could inhibit more than 70% LPS induced IL 8 expression ( P <0.05). Conclusion Transfection of A20 gene in HUVECs can inhibit the LPS induced activation of endothelial cells, which is helpful for the treatment of inflammation.