体外人血灌注猪肾异种肾移植超急性排斥反应试验探究.doc

体外人血灌注猪肾异种肾移植超急性排斥反应试验探究【摘要】 目的 体外人血灌注猪肾,初步探讨异种肾移植超急性排斥反应(hyperacute rejection,HAR)机理。方法 将猪肾随机分为对照组(1组,3例)采用猪自体血灌注,试验组分别采用人全血(2组,5例)、补体去活化人血(3组,5例)和去除天然抗体人血(4组,5例),灌注离体猪肾30 min,计算每5 min肾血流率和肾血管阻力,灌注结束后肾组织样本行免疫组化病理检查。结果 2组猪肾出现HAR,血管阻力增加,血流率下降,无泌尿。余三组猪肾未出现HAR,血管阻力及血流量维持稳定,持续有泌尿。结论 人血补体去活化或天然抗体廓清可避免灌注猪肾时的HAR。 【关键词】 异种移植;排斥反应;肾脏 Research of xenograft hyperacute rejection by perfusing pig kidneys with human blood in vitro XIONG Da-fu,JIA Ying-bin,GUAN Xiao-dong,et al. Department of General Surgery Ⅲ,the Fifth Affiliated Hospital of Sun Yat-Sen University,Zhuhai 519000, China 【Abstract】 Objective Ex vivo studies by perfusing pig kidneys with differents blood to evaluate human antiporcine xenograft hyperacute rejection.Methods Pig kidneys were perfused for 30 minutes with pig blood (group 1; n=3),human blood (group 2; n=5),complement heat innactivated human blood (group 3; n=5),and xenoreactive natural antibodies (XNA)-depleted human blood (group 4; n=5).Renal blood flow rate and vascular resistance were caculated very 5 minutes.Histologic and immunohistologic examinations were carried out after perfusion.Results Pig kidneys perfused with human blood,group 2,showed HAR,higher vascular resistance and lower blood flow rate.Kidneys in Group 1,3,4 had no histologic evidence of HAR.Blood flow rate and vasular resistance kept steady.Conclusion The depletion of complement or XNA in human blood protected pig kidneys form HAR. 【Key words】 Xenotransplant; Rejection; Kidney 逐年增多的等待肾移植患者使得肾脏的供需矛盾日益紧张,异种移植是缓解肾源短缺状况的可能途径之一。然而大量研究发现,人体内预存的反应性天然抗体(xenogenic natural antibody,XNA)可识别广泛存在于非灵长类动物血管内皮的a-1,3半乳糖(alpha 1,3-Galactose,a-Gal)抗原发生超急性排斥反应(hyperacute rejection,HAR),导致移植肾在数分钟之内丧失功能。克服HAR,首要的是了解其机制,本研究采用不同成分人血离体灌注猪肾,制造异种肾移植动物模型,初步探讨HAR病变机制。 1 资料与方法 1.1 一般资料 试验用猪共10只,购自中山大学北校区实验动物中心。取肾方法见笔者先前研究[1]。简单的说,气管插管全麻后,腹部“十”切口,腹主动脉插管,原位灌注HCA液,整块切取,在0℃～4℃生理盐水中修整。共20个肾脏,因不便于灌注,去除两个多支动脉肾脏。试验用猪肾共18个,质量(108±7)g,热缺血时间为0,冷缺血时间(32±5)min,随机分为四组,分别采用猪自体血(1组,n=3)、人全血(2组,n=5)、补体去活化人血(3