A20基因转导抑制异种移植中猪内皮细胞的活化

Inhibition of Porcine Aortic Endothelial Cells Activation by Transfection of A20 Gene cDNA

延缓性排斥反应 (DXR)是进行异种器官移植亟待解决的问题之一 .在DXR过程中 ,核心事件是异种移植物血管内皮细胞的活化 .核转录因子 (NF κB)在这一过程中起着重要的作用 .A2 0是一个具有锌指结构的蛋白 ,它可以抑制以NF κB为核心的信号转导系统 .向猪血管内皮细胞(PAEC)中导入A2 0基因 ,经RT PCR检测 ,A2 0基因能在细胞中稳定表达 .细胞生长曲线分析表明 ,A2 0的基因转导并不影响细胞的正常生长 ;电泳迁移率变动分析 (EMSA)表明 ,A2 0基因能抑制由肿瘤坏死因子 α(TNF α)诱导的NF κB的激活 ;流式细胞术分析表明 ,A2 0基因对受NF κB调控的一个重要炎症因子———E选择素的表达抑制率达 77 2 % .A2 0基因转导可能成为克服异种移植过程中DXR的手段之一 .

Activation of endothelial cells(EC) in xenografts is a key problem of delayed xenografts rejection (DXR) which is thought to be the main obstacle in xenotransplantation. Nuclear factor-κB plays an important role in DXR processes. A20, a novel zinc finger protein, is a critical negative regulator of TNF-induced NF-κB activation. Expression vector containing A20 cDNA was transferred into the porcine aortic endothelial cells (PAEC) PEDSV.15. The expression of A20 gene was stable in endothelial cells and had no effect on cell cycle. The results showed that expression of A20 gene in PAEC could inhibit the activation of NF-κB and reduce E-selectin expression by 77.2% detected by electrophoretic mobility shift assay (EMSA) and flow cytometry, respectively. All the data suggested that A20 gene transduction exhibited a useful method to overcome DXR in xenotransplantation.