E1A基因转导抑制小鼠血管内皮细胞的激活

Inhibition of Mouse Endothelial Activation by E1A Gene Microinjection

延缓性排斥反应 (delayedxenograftrejection ,DXR)是进行异种器官移植亟待解决的问题之一 .在DXR过程中 ,核心事件之一是核转录因子NF κB的激活 .人腺病毒 5 (Ad5 )的早期转录产物E1A蛋白可抑制以NF κB为核心的信号转导系统 .利用转基因技术向小鼠的受精卵导入了E1A基因 ,PCR和Southern印迹检测了 4 4只仔鼠 ,其中有 8只整合了E1A基因 .RT PCR检测发现 ,3只小鼠F1代的心、肝、肾等脏器都有E1AmRNA的表达 ,小鼠表型正常 .通过尾静脉向转基因阳性小鼠体内注射人灭活血清 ,模拟DXR发生的生理过程并用免疫荧光检测脏器细胞表面炎症分子E 选择素的表达水平 .结果表明 ,E1A基因的转导显著抑制了小鼠脏器细胞表面E 选择素的表达 。

NF κB activation is a key point in delayed xenograft rejection(DXR) which is thought to be the main obstacle in xenotransplantation. E1A, an early transcription protein of human adenovirus 5(Ad5), can inhibit the activation of NF κB. Transgenic vector containing E1A gene was constructed and 8 transgenic mice from 44 offspring were obtained by microinjection. RT PCR detection showed that F1 generation of 3 transgenic mice expressed the mRNA of E1A. Human serum was injected into F1 mice through caudal vein to induce the DXR. The expression of E selectin on the cell surface of organs was detected by immunofluorecence analysis. The results showed a remarkable reduction in the expression of E selectin in the liver, heart and kidney of transgenic mice. All the data suggested that E1A gene transfection was helpful in overcoming the DXR in transgenic mice.