白介素6基因敲除小鼠肝脏移植后的肝脏再生研究

目的 观察白介素6基因敲除(interleukin 6 knockout,IL-6 KO)小鼠原位肝移植后存活时间和肝脏再生的情况. 方法建立正常小鼠(C57BL/6 wild type,C57BL/6WT)和IL-6 KO小鼠的肝移植模型.38只小鼠分为3组:C57BL/6 WT→C57BL/6 WT肝脏移植对照组(n=10),IL-6 KO→IL-6 KO肝脏移植组(n=14)和IL-6 KO→C57BL/6 WT肝脏移植组(n=14).移植后观察小鼠肝移植物的存活情况.溴脱氧尿核苷(bromodeoxyuridine,BrdU)免疫组化检测移植肝脏的再生.结果 实验各组肝移植物的冷缺血时间均＜1 h.对照组小鼠肝移植后存活时间＞16 d.IL-6 K0→IL-6 KO组的肝移植后移植物不能存活(2 d).IL-6 KO→C57BL/6 WT组的肝移植后移植物不能存活(1.6 d).实验各组间存活时间比较,差异有统计学意义(F=190.09,P＜0.01).对照组小鼠肝脏移植后组织学检查证实肝脏组织损伤较轻,无组织坏死等改变.肝脏移植术后48 h BrdU摄取轻度增加.IL-6 KO小鼠肝移植后,组织学检查表现出片状坏死和肝细胞气球样变等改变.肝移植术后48 h免疫组化发现有极少数BrdU摄取.结论 IL-6 KO小鼠的肝移植后肝再生反应障碍,肝移植物不能存活.IL-6是肝脏再生反应中一个重要因子.     

MyD88及Trif对心脏移植小鼠体内供者特异性抗体及记忆性T淋巴细胞的影响

目的 探讨髓样分化因子88( MyD88)及Trif对皮肤移植预致敏的心脏移植小鼠体内供者特异性抗体(DSA)及记忆性T淋巴细胞的影响.方法 实验分为空白组(C57BL/6野生型小鼠,不行移植手术)、对照组(C3H小鼠为供者,C57BL/6小鼠为受者)和实验组(C3H小鼠为供者,MyD88及Trif基因敲除的C57BL/6小鼠为受者).将C3H小鼠皮片移植给受鼠,进行预致敏,2周后检测受鼠血清中 DSA的水平,并且将C3H小鼠心脏移植给相应受鼠,观察移植心脏存活时间.在观察终点时,再次检测受鼠血清中DSA水平,同时检测受鼠脾脏中记忆性T淋巴细胞的比例.结果 皮肤移植2周后,实验组受鼠DSA IgG2水平低于对照组,差异有统计学意义(P＜0.05),DSA IgG1和IgG3的水平亦低于对照组,但差异无统计学意义(P＞0.05).心脏移植3d后,与对照组相比较,实验组受鼠DSA IgG2水平明显降低(P＜0.01),受鼠脾脏中CD4+和CD8+记忆性T淋巴细胞的比例低于对照组(P＜0.01,P＜0.05).两组受鼠心脏移植后平均存活时间的差异无统计学意义(P＞0.05).结论 敲除小鼠MyD88及Trif基因虽然不能延长预致敏小鼠移植心脏的存活时间,但是能显著降低受鼠血清中DSA的水平及脾脏中记忆性T淋巴细胞的比例.     

CXC趋化因子受体6在小鼠心脏移植排斥反应中的表达及意义

目的 研究CXC趋化因子受体6(CXCR6)在同种异体小鼠心脏移植中的表达及CXC趋化因子配体16(CXCL16)与CXCR6相互作用对移植物存活时间的影响.方法 以野生型Balb/c小鼠(H-2d)为供者(同种移植组),或以野生型C57BL/6小鼠(H-2b)为供者(同系移植组),以野生型C57BL/6小鼠为受者分别行小鼠腹腔异位心脏移植.测定同系和同种移植组小鼠移植心脏CXCR6mRNA的表达,并测定受者脾脏CD8+T淋巴细胞CXCR6的表达.另制作小鼠同种异位心脏移植模型(Balb/c小鼠为供者,C57BL/6小鼠为受者),将其分为实验组和对照组,实验组受者移植当天至发生排斥反应时腹腔注射抗CXCL16抗体,对照组受者同期注射对照抗体.记录两组移植心脏存活时间.进行CD8+T淋巴细胞的细胞毒试验,即用Balb/c小鼠脾细胞免疫C57BL/6小鼠后,获取C57BL/6小鼠脾脏CD8+T淋巴细胞,将Balb/c小鼠脾细胞与C57BL/6小鼠CD8+T淋巴细胞混合培养,分别加入抗CXCL16抗体、小鼠IgG(对照抗体)和抗CD40L抗体.结果 同种移植组移植心脏中CXCR6 mRNA的表达以及脾脏CD8+T淋巴细胞上CXCR6的表达均高于同系移植组和正常对照组.抗CXCL16抗体对CD8+T淋巴细胞的细胞毒活性无影响.与对照组相比较,实验组小鼠移植心脏存活时间并未明显延长.结论 小鼠心脏移植排斥反应中CD8+T淋巴细胞CXCR6的表达上升,阻断CXCL16/CXCR6相互作用并不能延长移植心脏的存活时间.     

CXC趋化因子受体6在小鼠心脏移植排斥反应中的表达及意义

目的 研究CXC趋化因子受体6(CXCR6)在同种异体小鼠心脏移植中的表达及CXC趋化因子配体16(CXCL16)与CXCR6相互作用对移植物存活时间的影响.方法 以野生型Balb/c小鼠(H-2d)为供者(同种移植组),或以野生型C57BL/6小鼠(H-2b)为供者(同系移植组),以野生型C57BL/6小鼠为受者分别行小鼠腹腔异位心脏移植.测定同系和同种移植组小鼠移植心脏CXCR6mRNA的表达,并测定受者脾脏CD8+T淋巴细胞CXCR6的表达.另制作小鼠同种异位心脏移植模型(Balb/c小鼠为供者,C57BL/6小鼠为受者),将其分为实验组和对照组,实验组受者移植当天至发生排斥反应时腹腔注射抗CXCL16抗体,对照组受者同期注射对照抗体.记录两组移植心脏存活时间.进行CD8+T淋巴细胞的细胞毒试验,即用Balb/c小鼠脾细胞免疫C57BL/6小鼠后,获取C57BL/6小鼠脾脏CD8+T淋巴细胞,将Balb/c小鼠脾细胞与C57BL/6小鼠CD8+T淋巴细胞混合培养,分别加入抗CXCL16抗体、小鼠IgG(对照抗体)和抗CD40L抗体.结果 同种移植组移植心脏中CXCR6 mRNA的表达以及脾脏CD8+T淋巴细胞上CXCR6的表达均高于同系移植组和正常对照组.抗CXCL16抗体对CD8+T淋巴细胞的细胞毒活性无影响.与对照组相比较,实验组小鼠移植心脏存活时间并未明显延长.结论 小鼠心脏移植排斥反应中CD8+T淋巴细胞CXCR6的表达上升,阻断CXCL16/CXCR6相互作用并不能延长移植心脏的存活时间.     

CXC趋化因子受体6在小鼠心脏移植排斥反应中的表达及意义

目的 研究CXC趋化因子受体6(CXCR6)在同种异体小鼠心脏移植中的表达及CXC趋化因子配体16(CXCL16)与CXCR6相互作用对移植物存活时间的影响.方法 以野生型Balb/c小鼠(H-2d)为供者(同种移植组),或以野生型C57BL/6小鼠(H-2b)为供者(同系移植组),以野生型C57BL/6小鼠为受者分别行小鼠腹腔异位心脏移植.测定同系和同种移植组小鼠移植心脏CXCR6mRNA的表达,并测定受者脾脏CD8+T淋巴细胞CXCR6的表达.另制作小鼠同种异位心脏移植模型(Balb/c小鼠为供者,C57BL/6小鼠为受者),将其分为实验组和对照组,实验组受者移植当天至发生排斥反应时腹腔注射抗CXCL16抗体,对照组受者同期注射对照抗体.记录两组移植心脏存活时间.进行CD8+T淋巴细胞的细胞毒试验,即用Balb/c小鼠脾细胞免疫C57BL/6小鼠后,获取C57BL/6小鼠脾脏CD8+T淋巴细胞,将Balb/c小鼠脾细胞与C57BL/6小鼠CD8+T淋巴细胞混合培养,分别加入抗CXCL16抗体、小鼠IgG(对照抗体)和抗CD40L抗体.结果 同种移植组移植心脏中CXCR6 mRNA的表达以及脾脏CD8+T淋巴细胞上CXCR6的表达均高于同系移植组和正常对照组.抗CXCL16抗体对CD8+T淋巴细胞的细胞毒活性无影响.与对照组相比较,实验组小鼠移植心脏存活时间并未明显延长.结论 小鼠心脏移植排斥反应中CD8+T淋巴细胞CXCR6的表达上升,阻断CXCL16/CXCR6相互作用并不能延长移植心脏的存活时间.     

门静脉输注供者脾细胞诱导的供者特异性免疫低反应性

目的 探讨经门静脉输注供者脾细胞能否诱导皮肤移植小鼠产生供者特异性的免疫低反应性及其可能机制.方法 取Balb/c小鼠,随机分为空白对照组(经小鼠门静脉输注RPMI 1640培养液)、受者脾细胞组(经小鼠门静脉输注Balb/c小鼠脾细胞)、供者脾细胞组(经小鼠门静脉输注C57BL/6小鼠脾细胞)、空白移植对照组(经小鼠门静脉输注RPMI 1640培养液,7 d后移植C57BL/6小鼠的皮肤)、实验对照组(经小鼠门静脉输注Balb/c小鼠脾细胞,7 d后移植C57BL/6小鼠的皮肤)、实验组(经小鼠门静脉输注C57BL/6小鼠脾细胞,7 d后移植C57BL/6小鼠的皮肤)以及第三方移植组(经小鼠门静脉输注C57BL/6小鼠脾细胞,7 d后移植C3H小鼠的皮肤).记录空白移植对照组、实验对照组、实验组和第三方移植组移植皮肤的存活时间,并观察移植皮肤的病理学变化;脾细胞输注后7 d,分别获取空白对照组、受者脾细胞组和供者脾细胞组小鼠的外周血、脾脏和肝脏,用流式细胞仪测定样本中CD4+CD25+Foxp3+调节性T淋巴细胞(CD4+CD25+Foxp3+Treg细胞)的比例.结果 实验组移植皮肤的存活时间为(19.8±4.6)d,明显长于空白移植对照组、实验对照组和第三方移植组,但仍未达到长期存活.皮肤移植后7 d,空白移植对照组和实验对照组的移植皮肤呈现重度急性排斥反应的病理学改变,而实验组移植皮肤呈现中度急性排斥反应的病理学改变.供者脾细胞组外周血、肝脏和脾脏中CD4+CD25+Foxp3+Treg细胞比例明显高于空白对照组和受者脾细胞组.结论 门静脉输注供者脾细胞可特异性地延长供者皮肤移植物的存活时间,减轻移植物的排斥反应,该效应可能与受者体内的CD4+CD25+Foxp3+Treg细胞增加有关.     

输注转染FasL基因的供者树突状细胞减轻小鼠心脏移植排斥反应

目的 探讨转染FasL基因的供者树突状细胞(DC)对小鼠心脏移植排斥反应的影响.方法 分离并培养C57BL/6小鼠骨髓DC,然后采用脂质体法以自行构建的pTracer-FasL真核表达质粒转染DC.以C57BL/6小鼠为供者,Balb/c小鼠为受者,将其分为转染组(n=12)、未转染组(n=12)及移植对照组(n=12).转染组小鼠心脏移植前7 d经阴茎背静脉注射1×106个转染FasL基因的供者DC;未转染组小鼠心脏移植前7 d经阴茎背静脉注射1×106个未转染的供者DC;移植对照组仅行心脏移植,不接受供者DC输注.供心均移植于受者腹腔内.各组中,6只小鼠用于观察移植心脏存活时间,另6只于术后7 d处死,取移植心脏,进行组织学观察及移植心脏排斥反应病理分级.结果 转染组、未转染组和移植对照组小鼠移植心脏存活时间的中位数分别为20 d、8.5 d和9 d,转染组移植心脏的存活时间明显长于未转染组和移植对照组(P＜0.01).转染组中,2只排斥反应的病理分级为0级,4只为1级;未转染组中,2只为2级,4只为3级;移植对照组中,1只为2级,5只为3级.转染组排斥反应的病理分级明显低于移植对照组(P＜0.01).结论 受者于心脏移植前输注转染FasL基因的供者DC,可有效延长移植心脏的存活时间,并减轻排斥反应的程度.     

IL-23、IL-23 mRNA在皮肤移植受鼠外周血及移植物中的表达

目的 探讨皮肤移植小鼠外周血及移植物中白细胞介素23(IL-23)和IL-23 mRNA的表达情况.方法 制作小鼠皮肤移植的动物模型:同系组供、受者均为Balb/c小鼠;同种组以C57BL/6小鼠为供者,Balb/c小鼠为受者;脾细胞组以C57BL/6小鼠为供者,Balb/c小鼠为受者,受鼠移植前1 d经尾静脉输注供者脾细胞1×107个/只.移植术后1、3、5和7 d留取受者血液和皮肤移植物样本,检测血清IL-23的浓度以及移植皮肤中IL-23 mRNA的表达,并于光镜下持续观察移植皮肤的病理学变化.结果 移植后第3和5天,同系组受鼠外周血IL-23浓度分别为(263.49±18.72)和(286.55±18.95)pg/ml,同种组分别为(295.66±16.36)和(332.58±17.37)pg/ml,脾细胞组分别为(303.66±16.99)和(323.96±18.99)pg/ml.移植后第3和5天,同系组受鼠外周血IL-23浓度低于同种组和脾细胞组(P＜0.01);同种组和脾细胞组间的差异无统计学意义(P＞0.05).皮肤移植后第3、5和7天,同系组受鼠移植皮肤IL-23 mRNA的相对表达量分别为0.57±0.10、0.63±0.10和0.66±0.11,同种组分别为0.78±0.09、0.81±0.09和0.69±0.14,脾细胞组分别为0.62±0.10、0.68±0.12和0.55±0.09.移植后第3和5天,同系组和脾细胞组受鼠移植皮肤IL-23 mRNA的相对表达量低于同种组(P＜0.01).移植后第7天,同种组与脾细胞组受鼠移植皮肤IL-23 mRNA的相对表达量的差异有统计学意义(P＜0.01).结论 皮肤移植后早期发生急性排斥反应时,受鼠高表达IL-23和IL-23 mRNA.IL-23可作为预测排斥反应发生的观察指标.     

输注负载胎盘滋养层细胞抗原的受者aaDC延长小鼠移植心的存活时间

目的 探讨输注负载供者胎盘滋养层细胞抗原的受者耐受性选择性激活树突状细胞(aaDC)对小鼠移植心存活时间的影响.方法 雌性Balb/c小鼠和雄性C57BL/6小鼠合笼后,获取雌鼠胎盘滋养层细胞,并提取其抗原.体外制备Balb/c小鼠的aaDC,分别与滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原共培养,获得负载滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原的aaDC,测定培养液中白细胞介素10(IL-10)和IL-12的含量以及aaDC表面CD40、CD80、CD86和主要组织相容性复合物(MHC)Ⅱ类分子的表达.取Balb/c小鼠,分别接受负载滋养层细胞抗原或者C57BL/6小鼠脾细胞抗原的aaDC输注(滋养层细胞抗原组和脾细胞抗原组),7 d后接受C57BL/6小鼠心脏异位移植,以注射生理盐水者为对照组,输注负载滋养层细胞抗原aaDC后接受昆明小鼠心脏移植者为第三供者对照组.术后记录移植心存活时间,并检测受者脾脏和移植心组织中IL-2、IL-10和γ干扰素(IFN-7)mRNA的表达.结果 负载脾细胞抗原的aaDC,CD86和MHCⅡ类分子的表达明显升高,而负载滋养层细胞抗原的aaDC,仅MHCⅡ类分子表达升高.滋养层细胞抗原aalDC的培养液中,IL-10为(122.6±15.4)ng/L,IL-12为(232.8±25.4)ng/L,空白对照aaDC的IL-10和IL-12分别为(35.4±8.5)ng/L和(267.3±12.5)ng/L,二者间IL-10的差异有统计学意义(P<0.05),而IL-12的差异无统计学意义(P>0.05).脾细胞抗原aaDC培养液中的IL-10和IL-12与空白对照aaDC相近,差异均无统计学意义(P>0.05).对照组移植心存活时间为(6.73±0.58)d,脾细胞抗原组为(12.28±0.76)d,滋养层细胞抗原组为(38.83±2.79)d,第三供者对照组为(6.68±0.62)d.滋养层细胞抗原组移植心存活时间明显长于脾细胞抗原组和对照组(P<0.01),而第三供者对照组和对照组间移植心存活时间的差异无统计学意义(P>0.05).结论 预先输注负载供者胎盘滋养层细胞抗原的受者aaDC,可延长小鼠移植心的存活时间.     

CXC趋化因子受体6在小鼠心脏移植排斥反应中的表达及意义

目的 研究CXC趋化因子受体6(CXCR6)在同种异体小鼠心脏移植中的表达及CXC趋化因子配体16(CXCL16)与CXCR6相互作用对移植物存活时间的影响.方法 以野生型Balb/c小鼠(H-2d)为供者(同种移植组),或以野生型C57BL/6小鼠(H-2b)为供者(同系移植组),以野生型C57BL/6小鼠为受者分别行小鼠腹腔异位心脏移植.测定同系和同种移植组小鼠移植心脏CXCR6mRNA的表达,并测定受者脾脏CD8+T淋巴细胞CXCR6的表达.另制作小鼠同种异位心脏移植模型(Balb/c小鼠为供者,C57BL/6小鼠为受者),将其分为实验组和对照组,实验组受者移植当天至发生排斥反应时腹腔注射抗CXCL16抗体,对照组受者同期注射对照抗体.记录两组移植心脏存活时间.进行CD8+T淋巴细胞的细胞毒试验,即用Balb/c小鼠脾细胞免疫C57BL/6小鼠后,获取C57BL/6小鼠脾脏CD8+T淋巴细胞,将Balb/c小鼠脾细胞与C57BL/6小鼠CD8+T淋巴细胞混合培养,分别加入抗CXCL16抗体、小鼠IgG(对照抗体)和抗CD40L抗体.结果 同种移植组移植心脏中CXCR6 mRNA的表达以及脾脏CD8+T淋巴细胞上CXCR6的表达均高于同系移植组和正常对照组.抗CXCL16抗体对CD8+T淋巴细胞的细胞毒活性无影响.与对照组相比较,实验组小鼠移植心脏存活时间并未明显延长.结论 小鼠心脏移植排斥反应中CD8+T淋巴细胞CXCR6的表达上升,阻断CXCL16/CXCR6相互作用并不能延长移植心脏的存活时间.

Abstract：

Objective To investigate the expression of CXCR6 in allograft rejection and effect of CXCL16/CXCR6 interaction on allograft survival Methods Intra-abdominal heterotopic heart transplantation was performed using wild type (WT) Balb/c mice (H-2d) (allogeneic) as donors or WT C57BL/6 mice (B6, H-2b) (syngeneic) as donors, and using WT B6 mice as recipients. The intragraft expression of CXCR6 and expression of CXCR6 in CD8+ T cells of the spleens from syngeneic and allogeneic recipients were examined. The allogeneic recipients were further divided into the experimental group (n = 5) and control group (n = 6) randomly. The experiment group and control group were injected with anti-CXCL16 mAb or control mAb respectively until rejection occurred. The cardiac allograft survival in experimental group and control group was evaluated. Results Rejected allografts showed higher expression of CXCR6 than syngeneic cardiac grafts. More importantly,expression of CXCR6 in CD8+ T cells was also up-regulated by allograft rejection. However, injection of anti-CXCL16 mAb could not inhibit cytotoxic activity of CD8+ T cells. Moreover, experimental group could not prolong the cardiac graft survival time as compared with control group. Conclusion Expression of CXCR6 in CD8+ T cells is up-regulated in allograft rejection.     

IL-6强化的肠外营养对肝硬化大鼠肝切除术后残肝的影响

目的 探讨白细胞介素-6(IL-6)强化的肠外营养(PN)对肝硬化大鼠肝切除术后残肝的影响.方法 6只正常大鼠为正常对照组(A组),24只肝硬化大鼠随机分为4组(B～E组,n＝6);B组:肝硬化术前组,C组:肝切除+颈内静脉插管术后1 d组,D组:肝切除术后行PN 5 d组,E组:肝切除术后行PN+IL-6 5d组.测大鼠肝功能,炎症反应和脂质过氧化指标.肝组织ALB mRNA的表达.结果 与D组比较,E组血清AST、ALT、ALP显著下降(P＜0.05),血清ALB显著升高(P＜0.05);血清IFN-γ、TNF-α、MDA显著下降(P＜0.05),血清SOD活性显著升高(P＜0.05);肝组织ALB mRNA表达显著升高(P＜0.05).结论 PN+IL-6可以加快肝硬化大鼠肝切除术后肝功能的恢复,减轻炎症反应和脂质过氧化损伤,促进肝脏蛋白合成.     

The calcium channel blocker nisoldipine minimizes the release of tumor necrosis factor and interleukin-6 following rat liver transplantation

Kupffer cells, when activated, release toxic cytokines such as tumor necrosis factor (TNF), which can cause tissue injury. Takei et al. have reported that nisoldipine, a calcium channel blocker which decreases phagocytotic activity by Kupffer cells, also diminishes liver and lung injury and dramatically improves survival following liver transplantation [27]. Therefore, we studied the effect of nisoldipine on the time course of TNF and interleukin-6 (IL-6) release following cold storage and liver transplantation in the rat. Livers were stored under survival and non-survival conditions in cold Euro-Collins solution in the presence or absence of nisoldipine (1.4 µM). After storage, the effluent was collected for determination of cytokines. The liver was then transplanted orthotopically and serum was collected at various time intervals for up to 5 h. In the effluent, TNF levels were very low in both the control and nisoldipine-treated groups and IL-6 was not measurable. Furthermore, when livers were stored under survival conditions and transplanted (liver stored in the cold for 4 h), serum TNF (2 U/ml) and IL-6 (350 U/ml) values were minimal in both the control and nisoldipine-treated groups. In contrast, when livers were stored under non-survival conditions and transplanted (liver stored in the cold for 10 h), TNF levels increased to 15 ± 2 U/ml, 150 min after graft reperfusion, an increase which was prevented by nisoldipine (6.5 U/ml). Serum IL-6 levels were also elevated 300 min after transplantation in livers stored for 10 h. Nisoldipine also reduced the release of this cytokine. Serum transaminases (SGOT) were elevated to values around 2000 U/l 5 h following transplantation. In the nisoldipine-treated group, values were lower between 60 and 300 min. In the lung, interstitial and alveolar edema and cellular infiltration were detectable 5 h post-operatively and were diminished by nisoldipine. These data confirmed that TNF and IL-6 release were minimal following cold storage and transplantation of livers stored under survival conditions, but were elevated transiently after transplantation under non-survival conditions. Nisoldipine prevented cytokine release, most likely by blocking the activation of Kupffer cells, which may explain how it decreases liver and lung injury very early following liver transplantation.     

核因子-κB"圈套"策略对创伤性炎症大鼠肝脏炎症介质IL-6的作用研究

目的探讨核因子-κB（NF-κB）靶向性寡核苷酸（oligodeoxynucleotides,ODN）“圈套”策略对创伤性炎症大鼠肝脏炎症介质IL-6的影响.方法Wistar大鼠96只,随机分成生理盐水对照组、创伤性炎症组、“圈套”ODN组和变异“圈套”ODN组.运用凝胶阻滞实验检测创伤性炎症术后肝脏组织NF-κB的活性及合成“圈套”ODN的体外竞争抑制,用RT-PCR和ELISA法检测大鼠肝脏组织IL-6的mRNA水平和蛋白水平.结果大鼠创伤性炎症术后3 h肝脏NF-κB的活性开始升高,在术后12 h达高峰.肝脏组织IL-6 mRNA水平和蛋白水平也明显上升,与肝脏NF-κB的活性改变一致,“圈套”ODN在体外能有效地抑制NF-κB活性.“圈套”ODN治疗后大鼠肝脏组织IL-6的mRNA水平和蛋白水平均明显下降,肝脏功能明显好转,而变异“圈套”ODN却没有效果.结论NF-κB“圈套”策略通过特异性抑制NF-κB活性,可以有效地抑制创伤性炎症大鼠肝脏炎症介质IL-6的释放.     

Sewed revascularization for arterialized liver transplantation in mice

Background

Mouse models of liver transplantation are powerful tools for biomedical research. The cuff method is currently the most popular approach for revascularization of mouse liver grafts, as it is relatively easy to perform hence reducing the anhepatic time. However, the use of cuffs may induce a tissue reaction, causing chronic obstruction of anastomosed vessels, leading to portal hypertension. Here, we applied the suture technique for arterialized liver transplantation in mice.

Materials and methods

Liver transplantation was performed on 14 pairs of C57BL/6 mice. All hepatic vessels were anastomosed by sewing. The bile duct was connected with a stent. The liver grafts were harvested for histology on day 30 after surgery. Serum aspartate transaminase, alkaline phosphatase and bilirubin were measured at d 3, 7, and 30 after implantation.

Results

With a mean anhepatic time of 25.78 ± 3 min, the survival rate was 86% (n = 14) at 30 d following surgery. During this period, no significant liver injury was observed as assessed by serum markers and histology. Survival remained stable when grafts were exposed to 6 h cold ischemia prior to implantation. Vessel examination at the end of the studied period revealed an intact patency and a lack of collateral vessel growth.

Conclusion

Arterialized liver transplantation with sewed revascularization in mice is technically feasible. Both sewing and arterialization seem to be important factors promoting the survival of mouse recipients. The mouse model of suture arterialized orthotopic liver transplantation provides a novel tool for modern transplantation research and might be particularly suited for studies requiring longer-term survival of recipients.     

Systemic liberation of interleukin-8 in the perioperative phase of liver transplantation

Serum levels of interleukin-8 (IL-8) were investigated in the perioperative phase of liver transplantation (LTx) in order to help determine whether this cytokine might serve as a parameter for preservation injury. In a study of 45 patients undergoing LTx, systemic IL-8 was estimated at the end of the anhepatic phase, at 30, 60, and 120 min after reperfusion of the graft, and 24 h and 7 days after LTx. A maximum mean concentration of 665 ± 135 pg/ml was seen 60 min after LTx. The minimum was found on the 1st postoperative day (POD 1): 328 ± 33 pg/ml. Significant changes were found between 60 min and PODs 1 and 7, as well as between 120 min and POD 1. Differences in cold ischemia time were not found to be significant. We conclude that monitoring of systemic IL-8 levels is not useful in the development of new liver preservation concepts. Received: 12 September 1996 Received after revision: 27 February 1997 Accepted: 3 March 1997     

降钙素基因相关肽对急性肝损伤小鼠肝脏微循环的干预

目的：研究降钙素基因相关肽（CGRP）对刀豆蛋白A（ConA）诱导的小鼠肝损伤模型肝脏微循环的影响。方法：昆明种小鼠60只,随机分为ConA损伤组、CGRP干预组和空白对照组（n=20）,空白对照组注射生理盐水,ConA损伤组用ConA诱导建立小鼠急性肝损伤模型,CGRP干预组在用ConA诱导建模前体外注射CGRP,各组中10只于处理后用激光多普勒血流仪测肝脏的平均血流灌注量,另10只活体观察肝脏微循环流速,最后处死,全部HE染色观察肝脏病理变化。结果：CGRP干预组肝脏的平均血流灌注量和血液流速较ConA损伤组明显增加（P〈0.01）,病理学改变明显减轻。2组的血细胞浓度差异无统计学意义（P〉0.05）。结论：CGRP能够通过改变肝脏组织的灌注影响急性肝损伤时肝脏微循环障碍的程度和病理改变。     

输注外源性白细胞介素13延长同种肝移植大鼠的存活时间

目的 探讨外源性重组白细胞介素13(rIL-13)抑制同种肝移植大鼠的排斥反应和延长大鼠存活时间的作用及其机制.方法 以Lewis大鼠为供鼠,BN大鼠为受鼠,采用重建肝动脉的大鼠肝移植方法建立同种大鼠原位肝移植模型.采用随机数字表法将受鼠分为两组,实验组于术后第1、2、3、4和5天经尾静脉注射rIL-13 10μg/d,同期对照组注射等体积生理盐水.术后第7天,检测两组移植肝功能,测定移植肝组织病理改变、细胞因子表达及CD8+T淋巴细胞浸润等情况,并根据Banff标准计算排斥活动指数(RAI),观察术后2周的存活率.结果 与对照组相比,实验组肝功能明显改善,肝组织肿瘤坏死因子α(TNF-α)与E选择素的表达明显降低(P＜0.05),CD8+T淋巴细胞浸润明显减少(P＜0.05),术后2周存活率明显提高(P＜0.05).实验组的RAI为4.8±1.2,明显低于对照组的7.5±1.2(P＜0.05).结论 外源性重组rIL-13可减少促炎症因子TNF-α的释放和抑制E选择素的表达,减少移植物CD8+T淋巴细胞的浸润,从而减轻肝移植后急性排斥反应,延长大鼠的存活时间.     

Minimal increase in gut-mucosal interleukin-6 during laparoscopy

Background: The gut clearly plays a significant role in postoperative recovery. Other investigators have shown an increase in gut-mucosal cytokines in septicemia and burn models. We tested the effects of laparotomy and laparoscopy on gut-mucosal IL-6 production. Methods: A/J mice were randomized to three groups: control, laparotomy plus bowel manipulation (OBM), and laparoscopy plus bowel manipulation (LBM). Serum and gut-mucosal samples obtained at 4 and 8 h after surgery were analyzed for IL-6. Results: We found that OBM is associated with increased serum and gut-mucosal IL-6 at both 4 and 8 h after surgery. In contrast, LBM showed a blunted response in serum IL-6 and no change in gut-mucosal IL-6 at both time intervals. Conclusions: We conclude that laparoscopy minimizes trauma to the peritoneal environment, thereby decreasing the gut's inflammatory response to operation. This differential response of the gut may partially explain the preservation of gut function following laparoscopy.     

理想小鼠结直肠癌肝转移模型的建立及研究进展

结直肠癌是世界上第三大常见的恶性肿瘤,尽管对结直肠癌的发病机制、病理、预防和治疗方面的认识有了许多进展,但患者仍有较高的病死率,其病死率与肝脏转移密切相关,因此研究结直肠癌肝转移的机制和控制疾病进展是改善这些患者生存期的重要措施。近年来小鼠结直肠癌肝转移模型已广泛应用于转移研究,笔者就目前应用于结直肠癌肝转移的小鼠模型及其特点进行综述并分析它们的优缺点。以期为研究者在选择小鼠结直肠癌肝转移模型时提供参考,并为开发更为理想的小鼠结直肠癌肝转移模型指明方向,并为结直肠癌肝转移机制研究提供线索。