中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2008年
5期
257-260
,共4页
肝移植%大鼠%免疫耐受%白细胞介素%T细胞亚群
肝移植%大鼠%免疫耐受%白細胞介素%T細胞亞群
간이식%대서%면역내수%백세포개소%T세포아군
liver transplantation%rat%immune tolerance%interleukin%T lymphocyte subtype
目的 探讨白细胞介素-2(IL-2)mRNA及IL-10 mRNA表达水平对大鼠肝移植耐受的影响.方法 将实验大鼠随机分3组:Ⅰ组为急性排斥组; Ⅱ组为CD4+CD25+T细胞处理组,供体Wistar大鼠,受体SD大鼠;Ⅲ组为移植对照组,供体、受体均为SD大鼠.每组12对.肝移植前7 d,Ⅱ组受体大鼠经阴茎背静脉注射含供体脾淋巴细胞的培养液,Ⅰ组、Ⅲ组注射等量生理盐水;术后7 d随机取6只大鼠用逆转录-聚合酶链反应(RT-PCR)测定肝组织中细胞因子IL-2 mRNA及IL-10 mRNA的表达,用流式细胞仪检测各组移植肝内分离出的淋巴细胞含量,同时检测肝脏病理学的变化;另6只观察移植大鼠的生存期.结果 IL-2mRNA在Ⅰ组大鼠移植物内出现高表达,Ⅱ组仅有微弱表达,Ⅲ组则未见表达,IL-10 mRNA仅在Ⅱ组中表达,且表达程度较强.Ⅱ组和Ⅲ组大鼠存活期均超过30 d,与Ⅰ组(8～11 d)比较差异有统计学意义(P均＜0.01).Ⅰ组大鼠移植后肝脏有大量淋巴细胞浸润,数量明显高于Ⅱ组和Ⅲ组[(14.31±3.41)×106 /g比(5.04±1.13)×106 /g和(1.55±0.40)×106 /g,P均＜0.01],且显示中度排斥反应.Ⅱ组大鼠有中等量淋巴细胞浸润,病理为无排斥或不确定性排斥,且淋巴细胞中CD4+百分比[(43.31±8.07)%]和CD4+CD25+百分比C(11.39±1.92)%]均显著高于Ⅰ组[(33.65±7.25)%,(3.05±0.62)%]和Ⅲ组[(31.18±6.52)%,(3.37±0.72)%],差异有统计学意义(P＜0.05或P＜0.01).Ⅲ组未见明显淋巴细胞浸润,病理为无排斥反应.结论 IL-2参与了移植排斥的发生,而IL-10在CD4+CD25+T细胞诱导免疫耐受中具有非常重要的作用.
目的 探討白細胞介素-2(IL-2)mRNA及IL-10 mRNA錶達水平對大鼠肝移植耐受的影響.方法 將實驗大鼠隨機分3組:Ⅰ組為急性排斥組; Ⅱ組為CD4+CD25+T細胞處理組,供體Wistar大鼠,受體SD大鼠;Ⅲ組為移植對照組,供體、受體均為SD大鼠.每組12對.肝移植前7 d,Ⅱ組受體大鼠經陰莖揹靜脈註射含供體脾淋巴細胞的培養液,Ⅰ組、Ⅲ組註射等量生理鹽水;術後7 d隨機取6隻大鼠用逆轉錄-聚閤酶鏈反應(RT-PCR)測定肝組織中細胞因子IL-2 mRNA及IL-10 mRNA的錶達,用流式細胞儀檢測各組移植肝內分離齣的淋巴細胞含量,同時檢測肝髒病理學的變化;另6隻觀察移植大鼠的生存期.結果 IL-2mRNA在Ⅰ組大鼠移植物內齣現高錶達,Ⅱ組僅有微弱錶達,Ⅲ組則未見錶達,IL-10 mRNA僅在Ⅱ組中錶達,且錶達程度較彊.Ⅱ組和Ⅲ組大鼠存活期均超過30 d,與Ⅰ組(8～11 d)比較差異有統計學意義(P均＜0.01).Ⅰ組大鼠移植後肝髒有大量淋巴細胞浸潤,數量明顯高于Ⅱ組和Ⅲ組[(14.31±3.41)×106 /g比(5.04±1.13)×106 /g和(1.55±0.40)×106 /g,P均＜0.01],且顯示中度排斥反應.Ⅱ組大鼠有中等量淋巴細胞浸潤,病理為無排斥或不確定性排斥,且淋巴細胞中CD4+百分比[(43.31±8.07)%]和CD4+CD25+百分比C(11.39±1.92)%]均顯著高于Ⅰ組[(33.65±7.25)%,(3.05±0.62)%]和Ⅲ組[(31.18±6.52)%,(3.37±0.72)%],差異有統計學意義(P＜0.05或P＜0.01).Ⅲ組未見明顯淋巴細胞浸潤,病理為無排斥反應.結論 IL-2參與瞭移植排斥的髮生,而IL-10在CD4+CD25+T細胞誘導免疫耐受中具有非常重要的作用.
목적 탐토백세포개소-2(IL-2)mRNA급IL-10 mRNA표체수평대대서간이식내수적영향.방법 장실험대서수궤분3조:Ⅰ조위급성배척조; Ⅱ조위CD4+CD25+T세포처리조,공체Wistar대서,수체SD대서;Ⅲ조위이식대조조,공체、수체균위SD대서.매조12대.간이식전7 d,Ⅱ조수체대서경음경배정맥주사함공체비림파세포적배양액,Ⅰ조、Ⅲ조주사등량생리염수;술후7 d수궤취6지대서용역전록-취합매련반응(RT-PCR)측정간조직중세포인자IL-2 mRNA급IL-10 mRNA적표체,용류식세포의검측각조이식간내분리출적림파세포함량,동시검측간장병이학적변화;령6지관찰이식대서적생존기.결과 IL-2mRNA재Ⅰ조대서이식물내출현고표체,Ⅱ조부유미약표체,Ⅲ조칙미견표체,IL-10 mRNA부재Ⅱ조중표체,차표체정도교강.Ⅱ조화Ⅲ조대서존활기균초과30 d,여Ⅰ조(8～11 d)비교차이유통계학의의(P균＜0.01).Ⅰ조대서이식후간장유대량림파세포침윤,수량명현고우Ⅱ조화Ⅲ조[(14.31±3.41)×106 /g비(5.04±1.13)×106 /g화(1.55±0.40)×106 /g,P균＜0.01],차현시중도배척반응.Ⅱ조대서유중등량림파세포침윤,병리위무배척혹불학정성배척,차림파세포중CD4+백분비[(43.31±8.07)%]화CD4+CD25+백분비C(11.39±1.92)%]균현저고우Ⅰ조[(33.65±7.25)%,(3.05±0.62)%]화Ⅲ조[(31.18±6.52)%,(3.37±0.72)%],차이유통계학의의(P＜0.05혹P＜0.01).Ⅲ조미견명현림파세포침윤,병리위무배척반응.결론 IL-2삼여료이식배척적발생,이IL-10재CD4+CD25+T세포유도면역내수중구유비상중요적작용.
Objective To investigate the expression of tolerance-associated interleukin (IL)-2 mRNA and IL-10 mRNA in rats after allogeneic liver transplantation. Methods The experimental rats were randomly divided into 3 groups : acute rejection model group (group Ⅰ ), CD4+CD25+T cell treatment group (group Ⅱ), donors of the two groups were Wistar while recipients were Spregue-Dawley (SD) rats, and group Ⅲ as control group, both donors and recipients of this group were SD rats, with 12 rats in each group.Splenic lymphocyte of donor in group Ⅱ were injected through vena dorsalis penis 7 days before liver transplantation; equal volume normal saline (NS) were injected in rats of group Ⅰ and Ⅲ. The contents of IL-2 mRNA and IL-10 mRNA in the liver tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR), and the number of lymphocytes in the donor liver was quantified with flow cytometry 7 days after transplantation. Meanwhile the pathologic change in the donor tissue were observed, and the recipients' life span was recorded. Results IL-2 mRNA was expressed strongly in the liver of group Ⅰ , but it was expressed weakly in group Ⅱ , and no expression was detected in the liver of group Ⅲ. IL-10 mRNA was expressed only in group Ⅱ , and the level was high. The rats in group Ⅱ and Ⅲ survived over 30 days,but rats in Ⅰ group had a shorter life (8 - 11 days, both P＜0. 01). There was heavy lymphocytic infiltration in the liver of group Ⅰ ,and was much higher than that of the other groups [group Ⅰ : (14.31±3.41)×106 /g,group Ⅱ : (5.04±1.13)×106 /g, group Ⅲ : (1.55±0. 40)×106 /g, all P＜0. 01], and pathology showed moderate rejection. Rats in group Ⅱ had milder lymphocytic infiltration, and pathology showed no signs of rejection or uncertain rejection, and the ratio of CD4+ T cell and CD4+CD25+ were higher than those of group Ⅰ [(43. 31±8. 07)% vs. (33.65±7.25)% and (11.39±1.92)% vs. (3.05±0.62)%] and group Ⅲ [(43.31±8. 07)% vs. (31.18±6.25)% and (11.39±1.92)% vs. (3.37±0. 72)%, P＜0. 05 or P＜0. 01].In group Ⅲ no lymphocytic infiltration was found, and pathology showed no sign of rejection. Conclusion IL-2 may participate in the immune rejection in allogeneic liver transplantation, but IL-10 plays an important role in CD4+CD25+T cell inducing immune tolerance of allogeneic liver transplantation in the rats.
