锌指蛋白基因对内毒素诱导选择素表达的影响

目的　探讨外源性A2 0基因对内毒素诱导的内皮细胞E 选择素表达的影响。　方法　DOTAP脂质体介导的pCNDA3 1EHA2 0基因转染人脐静脉内皮细胞 ,经G4 18筛选阳性克隆 ,免疫荧光检测A2 0基因的表达 ,原位杂交、免疫荧光及Westernblot分别检测内皮细胞E 选择素的表达。　结果　A2 0基因在经G4 18筛选后的内皮细胞中有高表达 ;内毒素能诱导人内皮细胞E 选择素表达 ;A2 0基因能抑制内毒素诱导的内皮细胞 90 %的E 选择素表达 ,两者间相差显著 (P <0 0 5 )。　结论　A2 0基因能够显著抑制内毒素诱导的内皮细胞活化 ,有助于创伤的治疗。

OVEREXPRESSION OF ZINC FINGER PROTEIN INHIBITS LPS-INDUCED EXPRESSION OF E-SELECTIN IN ENDOTHELIAL CELLS

Objective Previous studies have demonstrated that LPS can induce endothelial cell activation and the expression of E-selectin. In this study, we examined whether A20 gene could inhibit the expression of E-selectin in endothelial cells induced by LPS. Methods With the help of DOTAP, endothelial cells were transfected with pCDNA3.1 EHA20. The postive 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 immunofluorescence, Western blot and in situ hybridization. Results Abundant A20 stable expression in endothelial cells transfected with pCDNA3.1 EHA20 was confirmed by immunofluorescence analysis. E-selectin expression increased in LPS-inducible endothelial cells. A20 gene inhibited 90% LPS-inducible E-selectin expression(P<0.05). Conclusion With the help of DOTAP,hA20 gene can be transferred and stably expressed in endothelial cells and it can inhibit LPS-induced E-selectin expression of endothelial cells.