摘要
目的 观察腺相关病毒（AAV）介导的程序性死亡配体-1（PD-L1）基因转染对大鼠肝移植术后干扰素（INF）-γ、白细胞介素（IL）-17、IL-10及转化生长因子-β1 （TGF-β1）等的影响.方法 96只近交系大鼠随机分为等基因组（G1）、同种异体组（G2）、空载体组（G3）和转染组（G4）,每组12对.通过大鼠肝移植急性排斥模型,采用夹闭法将PD-L1-AAV2-红色荧光蛋白（RFP）转染供肝,观察大鼠中位生存时间（MST）,全自动生化分析仪测定肝功能,采用酶联免疫吸附试验（ELISA）法检测受体移植肝IL-10、IL-17、INF-γ及TGF-β1的表达.结果 G1组全部存活;G2、G3组MST分别为18（17 ～20）、18（17～19） d;G4组显著延长了其存活（MST 29 d）（P＜0.05）.术后14d G4组肝功能及病理变化与7d G2、G3组相似.除G1组1NF-γ无表达外,余组移植肝INF-γ与IL-17表达类似.G1组术后7 d IL-17水平明显升高,后渐降至28.9 ng/g,仍高于3d;G2、G3组移植肝INF-γ、IL-17水平3d高表达,并渐增强,均在14d达峰值418.6、382.9 ng/g.而G4组虽高表达,但其幅度及浓度均低于G2、G3组,除G4组3d外,差异有统计学意义（Jp＜0.05）.各组移植肝TGF-β1与IL-I0表达类似,G1组术后3d升高至170.2、106.2 ng/g,后渐降,但仍高于G2、G3组;而G4组术后3d高表达,14d达峰值564.1、614.9 ng/g,除G4组3d外,各组TGF-β1、IL-10均高于G2、G3组,差异有统计学意义（P＜0.05）.结论 AAV介导的PD-L1基因转染可延长移植肝的存活,其机制可能是基因转染增强了PD-L1/PD-1信号通路,在上调IL-10、TGF-β1表达的同时,下调INF-γ、IL-17的表达,从而发挥对移植肝的保护作用.
Objective To explore the adeno-associated virus-mediated （AAV） programmed death ligand-1 （PD-L1） gene transfection effect in rats after liver transplantation interferon （INF）-γ,interleukin （IL）-17,IL-10 and transforming growth factor-β1 （TGF-β1）.Methods All rats were divided into isogenic liver transplantation group （G1）,allogeneic liver transplantation group （G2）,allogeneic liver transplantation group infected by AAV2-RFP （Red fluorescent protein,RFP） （G3） and PD-L1-AAV2-RFP （G4）.AAV mediated PD-L1 gene transfer was performed in LEW rat donor livers placed in BN recipients.Median surv ival time （MST） of grafts were investigated.The function of liver was measured with automatic biochemical analyser.The expressions of IL-10,IL-17,INF-γ and TGF-[β1 in liver were assessed by Enzyme linked immunosorbent assay （ELISA）.Results Liver allograft survival was significantly prolonged by PD-L1-AAV2-RFP treatment [MST,29 （range：28-31） days] compared with either G2 or G3：18 （17-20） and 18 （17-19） days respectively （P＜0.05）.In addition to G1 no expression of INF-γ,the remaining groups of transplanted liver INF-γ was similar to that of IL-17.G1 after 7 d IL-17 significantly increased,then decreased gradually to 28.9 ng/g,still higher than the first 3 d.G2 and G3 graft INF-γ,IL-17 has a strong start from the first 3 d expression gradually increased,both in 14 d reached the peak value 418.6,382.9 ng/g.Although G4 expression increased,its range and concentrations were significantly lower than G2 and G3.In addition G4 3 d,the difference was significant.Each group transplanted liver TGF-β1 and IL-10 expression characteristics similar,G1 significantly increased when postoperative 3 d to 170.2,106.2 ng/g,gradually decreased,but still higher than the G2 and G3.The G4 3 d its expression began to increase after the first peak 14 d which reached the value of 564.1,614.9 ng/g.Each group TGF-β1 and IL-10 concentrations were significantly higher rejection group except G4 3 d （P ＜ 0.05）.Conclusion AAV-mediated gene transfer of PD-L1 into cold preserved liver allograft before transplantation results in long term survival of liver allograft.PD-L1 gene may take the effect in liver transplantation via suppression of INF-γ and IL-17 production as well as stimulation of IL-10 and TGF-β1 production.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第12期2583-2586,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30400418）
