锌指蛋白A20在内毒素所致人脐静脉内皮细胞损伤中的作用

OVEREXPRESSION OF A20 INHIBITS LPS-INDUCIBLE ENDOTHELIAL CELLS INJURY

目的 探讨外源性锌指蛋白A2 0对内毒素 (LPS)诱导的内皮细胞同型半胱氨酸蛋白酶caspase 3表达和凋亡的影响。 方法 RT PCR检测A2 0基因在内毒素诱导内皮细胞中的表达。DOTAP脂质体转染pCDNA3 1EHA2 0于脐静脉内皮细胞 ,经G4 18筛选 ,A2 0表达经免疫荧光鉴定。Tunnel原位末端标记法、原位杂交检测转染前后内毒素诱导内皮细胞凋亡情况及caspase 3的表达情况。 结果 A2 0基因在内毒素诱导的内皮细胞中能表达。正常对照组及转染A2 0基因组人脐静脉内皮细胞 (HUVEC)凋亡为 (5± 1) %、(6± 1) % ,LPS作用后对照组与转染A2 0基因组凋亡分别为 (36± 3) %、(10± 1) % ,两者相差显著 (P <0 0 5 )。A2 0基因能显著抑制内毒素诱导的内皮细胞caspase 3的表达。 结论 A2 0基因在创伤的治疗中可能有一定作用。

Objective To examine the effect of A20 on LPS inducible apoptosis and caspase-3 expression of human umbilical vein endothelial cells. Methods Using DOTAP the endothelial cells were transfected with pCDNA3 1EHA20. positive cell clones were selected with G418. The stable transfection and expression of A20 in the endothelial cells were determined by immunofluorescence analysis. The expression of the A20 gene induction by LPS was checked by RT PCR. The caspase-3 expression was checked by in situ hybridization, apoptosis of the endothelial cells was observed with flow cytometry. Results The A20 gene was able to express in the endothelial cells induced by LPS. The A20 gene transfected by DOTAP was able to inhibit 90% LPS inducible caspase-3 expression and apoptosis of the endothelial cells(P<0 05).Conclusion A20 gene may be helpful for the treatment of trauma.