致敏大鼠肝移植后抗体介导的排斥反应的模型研究  

Study of the rejection mediated by antibodies on the transplantation model of sensitized rat

作　　者：赵姣姣 张升宁 莽源祎 高杨 李立 ZHAO Jiaojiao;ZHANG Shengning;MANG Yuanyi;GAO Yang;LI Li(Hepatobiliary Pancreaticovascular Surgery and Liver Transplantation Center,Calmete Hospital Affiliated to Kunming Medical University/The First Hospital of Kunming,Yunnan Provincial Organ Transplantation Clinical Medical Center,Kunming 65001l,P.R.China)

机构地区：[1]昆明医科大学附属甘美医院(昆明市第一人民医院)肝胆胰血管外科暨肝移植中心、云南省器官移植临床医学中心,昆明650011

出　　处：《中国普外基础与临床杂志》2023年第6期674-680,共7页Chinese Journal of Bases and Clinics In General Surgery

基　　金：云南省器官移植临床医学中心资助项目(项目编号:ZX2019-07-01);云南省科技厅科技计划项目(项目编号:202201AY070001-198)。

摘　　要：目的建立同种异体致敏大鼠原位肝移植模型,研究该模型发生的排斥反应是否符合抗体介导的排斥反应(antibody-mediated rejection,AMR)的病理学特点。方法以12只雄性Lewis大鼠为受者,250~290 g,6只雄性Brown Norway(BN)大鼠为供者,250~280 g。将Lewis大鼠采用随机数字表法随机分为4组(n=3);Lewis对照组(LC组,未做任何处理)、直接移植组(T组,BN大鼠的肝脏直接移植于Lewis大鼠体内)、致敏组(S组,Lewis大鼠体内输入BN大鼠的脾脏淋巴细胞)和致敏后移植组(TS组,Lewis大鼠体内输入BN大鼠脾脏淋巴细胞2周后,进行肝移植)。于肝移植后大鼠存活第14天,收集受者非致死量血液3~4 mL,检测天冬氨酸转氨酶(aspartate aminotransferase,AST)、丙氨酸转氨酶(alanine aminotransferase,ALT)、总胆红素(total bilirubin,TB)、碱性磷酸酶(alkaline phosphatase,ALP)和肌酐(creatinine,CRE)水平,并检测Lewis大鼠体内供者特异性抗体(donor-specific alloantibody,DSA)及补体C4d表达情况。采用苏木精-伊红染色和Masson染色评定4组大鼠肝脏组织的形态学指标,采用CK-19、C4d和CD20免疫组织化学染色方法评定4组大鼠肝脏发生排斥反应的程度并进行排斥反应活性指数(rejection activity index,RAI)评分。结果与T组相比,TS组Lewis大鼠的血清AST、TB和ALP水平以及DSA(IgG1、IgG2a、IgG2b、IgG2c)和C4d表达阳性率升高。与LC组相比,T组的Lewis大鼠出现部分胆管水肿及淋巴细胞浸润,但未见明显毛细血管结构的破坏;与T组相比,TS组Lewis大鼠的胆管前毛细血管出现大量的淋巴细胞或单核细胞浸润及毛细血管的严重破坏;TS组的RAI评分和C4d评分均高于T组。结论致敏大鼠肝移植后肝脏发生更严重的急性排斥反应和移植肝失功能,且致敏大鼠的急性排斥反应符合AMR的特点。由于本研究样本量较少,对AMR模型的探索还有待深入。Objective To explore the pathological features of rejection reaction and whether it accord with antibody-mediated rejection(AMR)in the liver transplantation model of allo-sensitized rat.Methods Twelve male Lewis rats as the recipient,250–290 g;6 male Brown Norway(BN)rats as the donor,250–280 g.Twelve Lewis recipient rats were randomly divided into 4 groups by random number method(n=3):Lewis control group(LC group,without any treatment),direct transplantation group(T group,livers from BN rats were directly transplanted into Lewis rats),sensitized group(S group,spleen lymphocytes from BN rats were injected into Lewis rats),and sensitized transplantation group(TS group,splenic lymphocytes from BN rats were injected into Lewis rats for 2 weeks before liver transplantation).On the 14th day after liver transplantation,3–4 mL of recipient non-lethal blood was collected to detect serum aspartate aminotransferase(AST),alanine aminotransferase(ALT),total bilirubin(TB),alkaline phosphatase(ALP)and creatinine(CRE)levels,and detect the expression of donor-specific alloantibody(DSA)and complement C4d in recipients.Hematoxylin-eosin(HE)staining and Masson staining were used to evaluate the morphological indexes of rat liver tissue,and CK-19,C4d and CD20 immunohistochemical staining methods were used to evaluate the degree of liver rejection and rejection activity index(RAI)score was performed.Results Compared with the T group,the serum AST,TB,and ALP levels,as well as the positive rates of DSA(IgG1,IgG2a,IgG2b,IgG2c)and C4d expression in Lewis rats in the TS group increased.Compared with the LC group,rats in the T group showed partial bile duct edema and lymphocyte infiltration,but no obvious damage of capillary structure was observed.Compared with the T group,a large number of lymphocytes or monocytes were infiltrated and capillaries were severely damaged in the anterior bile duct of rats in the TS group.The RAI and C4d scores of the TS group were higher than those of the T group.Conclusions More severe acute rejection a

关 键 词：供者特异性抗体 抗体介导的排斥反应 肝移植 大鼠 

分 类 号：R657.3[医药卫生—外科学]