骨髓来源未成熟树突状细胞干预大鼠原位肝移植的排斥反应  

Effect of immature dendritic cells derived from bone marrow on rejection of orthotopic liver transplantation in rats

作　　者：李立强 焦龙杏 张武 闫文涛 李健 李明皓[4] Li Liqiang;Jiao Longxing;Zhang Wu;Yan Wentao;Li Jian;Li Minghao(Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Wuzhong People’s Hospital,Wuzhong 751100,Ningxia Hui Autonomous Region,China;Hefei Changrong Hospital,Hefei 230001,Anhui Province,China;Department of Hepatobiliary Surgery,People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学,宁夏回族自治区银川市750004 [2]吴忠市人民医院,宁夏回族自治区吴忠市751100 [3]合肥长荣医院,安徽省合肥市230001 [4]宁夏回族自治区人民医院肝胆外科,宁夏回族自治区银川市750002

出　　处：《中国组织工程研究》2021年第13期2025-2029,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区科学基金项目(81560114),项目负责人:李明皓;宁夏重点研发项目(2019BEG03039),项目负责人:李明皓。

摘　　要：背景:未成熟树突状细胞具有很强的抗原摄取加工能力,但由于缺乏多种共刺激分子不能使初始T细胞活化、增殖产生免疫应答,可导致T细胞无能,从而诱导免疫低反应或抗原免疫特异性耐受,同时未成熟树突状细胞可诱导同种异体抗原特异性T细胞的低反应性,从而延长移植器官的存活时间。目的:探讨未成熟树突状细胞对大鼠原位肝移植术后肝脏排斥反应的影响及相关机制。方法:根据DA和Lewis大鼠体质量成对随机分配成3组,均利用“二袖套”方法建立DA→Lewis大鼠肝移植模型。对照组不给予任何处理;环孢素组术后第2天开始给予10 mg/kg环孢素A,1次/d,共7 d;未成熟树突状细胞组术前1 d经阴茎背静脉注射DA大鼠骨髓来源未成熟树突状细胞(1×10^6/只),连续注射2次,间隔10 min。术后7 d取大鼠肝脏,苏木精-伊红染色观察病理改变,qRT-PCR和Western blot检测SHIP、AKT、IKK、IKβmRNA以及蛋白的表达。结果与结论:①与对照组相比较,环孢素组和未成熟树突状细胞组生存时间较长,差异有显著性意义(P<0.05);②环孢素组和未成熟树突状细胞组大鼠肝组织汇管区单核细胞和淋巴细胞浸润较少,肝小叶结构未被明显破坏,肝动脉、门静脉和胆管的炎性细胞明显少于对照组,未达到重急性排斥反应水平;③对照组比较,环孢素组和未成熟树突状细胞组IKβmRNA表达升高,SHIP、AKT、IKK mRNA表达下降,差异均有显著性意义(P<0.05);④与对照组比较,未成熟树突状细胞组SHIP、IKβ蛋白表达升高,IKK、AKT蛋白表达下降,差异有显著性意义(P<0.05);与对照组比较,环孢素组SHIP、IKK蛋白表达下降,AKT、IKβ蛋白表达升高,差异有显著性意义(P<0.05);⑤结果表明,未成熟树突状细胞有减缓急性排斥反应的功能,延迟肝脏成活时间,降低同种异体肝脏移植的T细胞免疫应答能力。BACKGROUND:Immature dendritic cells have strong antigen uptake and processing ability,but the lack of a variety of costimulatory molecules cannot activate and proliferate initial T cells to produce immune response,which can lead to T cell anergy,thus inducing low immune response or antigen immune specific tolerance.Simultaneously,immature dendritic cells can induce hyporeactivity of allogeneic antigen-specific T cells,thus prolonging the survival time of transplanted organs.OBJECTIVE:To investigate the effect of immature dendritic cells on liver rejection after orthotopic liver transplantation in rats and its mechanism.METHODS:According to the weight of DA and Lewis rats,the rats were randomly divided into three groups,and the liver transplantation model of DA-Lewis rats was established by“two-cuff”method.The rats of control group received no measures.The rats of cyclosporine group were treated with 10 mg/kg cyclosporine from the second day after operation,once a day,for 7 days.The rats of the immature dendritic cell group were injected with 1×10^6 immature dendritic cells from bone marrow of DA rats one day before operation through dorsal penile vein;the injection was repeated twice with an interval of 10 minutes.The livers of all these rats were removed 7 days after operation.Hematoxylin-eosin staining was used to observe the pathological changes.The mRNA and protein expressions of SHIP,AKT,IKK and IKβin these three groups were detected by qRT-PCR and western blot assay.RESULTS AND CONCLUSION:(1)Compared with the control group,the survival time of cyclosporine group and immature dendritic cell group was significantly longer(P<0.05).(2)In cyclosporine group and immature dendritic cell group,the number of infiltrating mononuclear cells and lymphocytes in the portal area of liver tissue was less,the structure of hepatic lobule was not significantly damaged,and the inflammatory cells in hepatic artery,portal vein and bile duct were significantly less than those in control group,which did not reach the level o

关 键 词：肝移植 免疫排斥 未成熟树突状细胞 排斥反应 T细胞 免疫应答 大鼠 

分 类 号：R459.9[医药卫生—治疗学] R392-3[医药卫生—临床医学]