趋化因子CXC家族成员在大鼠肝移植急性排斥反应中的作用

目的探讨趋化因子Mig、IP-10、I-TAC及其受体CXCR3的表达在大鼠肝移植手术前后的动态变化,分析其与肝移植急性排斥（AR）的关系。方法改良“二袖套”法建立大鼠原位肝移植模型和AR模型,分为4组：手术创伤组,肝移植无排斥组,肝移植急性排斥组,免疫抑制剂组。EHSA法检测血清Mig、IP-10、I-TAC表达。流式细胞仪检测外周血淋巴细胞CXCR3的表达,用Cellquest软件分析阳性细胞百分率。半定量RT—PCR检测各组第7天肝脏组织CXCR3-mRNA的表达。结果（1）各组大鼠手术后外周血趋化因子Mig,IP-10,I-TAC及其受体CXCR3表达出现明显升高。（2）AR组在术后第3～5天起Mig,IP．10,I-TAC及其受体CXCR3表达均高于其他各组（P〈0．01）。（3）AR组大鼠在移植后第7天,外周血Mig、IP-10、I—TAC和CXCR3的表达随着AR组RAI积分的升高而逐步升高,相关系数分别为0．79,0．89,0．82,0．92（P〈0．05）。结论血清中趋化因子Mig、IP-10、I-TAC及其受体CXCR3的高表达与AR密切相关,有望成为诊断AR较特异、敏感的指标。     

趋化因子受体CXCR3动态检测在早期诊断肝移植术后急性排斥中的应用价值

目的：探讨趋化因子受体 CXCR3动态变化与肝移植排斥的关系及其在早期诊断中的意义。方法：以 2005年4月～2005年9月在本院行肝移植术的30例病人为研究对象，采用流式细胞仪检测病人术前1d 及术后第 1、 3、 5、 7天外周血中淋巴细胞 CXCR3的表达，临床诊断为急性排斥的病人在确诊当天以及经激素冲击治疗后第3、 7天各检测病人外周血中淋巴细胞 CXCR3的表达变化。结果：急性排斥反应(AR)组病人的 CXCR3表达均明显升高， 与非排斥组、肝癌肝硬化组及正常人对照组有明显差异(P＜0． 01) ，当 AR 组经冲击治疗逆转后 CXCR3表达明显下降 并维持持较低水平。结论：外周血中 CXCR3的表达与排斥反应密切相关，可作为诊断急性排斥反应的发生及观察抗 排异疗效的指标。     

反义肽核酸阻断CXCR3表达延长大鼠肝脏移植物存活期

目的研究反义肽核酸（antisense peptide nucleic acid,asPNA）阻断CXC趋化因子受体3（CXCR3）表达延长大鼠肝脏移植物存活期的作用和机制,探讨asPNA技术治疗肝移植急性排斥反应（acute rejection,AR）的潜在临床价值。方法采用改良＂二袖套＂法建立大鼠原位肝移植模型。分为4组：肝移植急性排斥组,肝移植免疫抑制剂组,asPNA组,asPNA对照组。RT-PCR法检测T细胞内CXCR3-mRNA表达水平,Western-blotting免疫印迹法检测CXCR3蛋白表达水平,流式细胞仪测定外周血CD3＋T细胞表面CXCR3表达水平。AR诊断参照Banff标准。结果asPNA在体外能有效抑制T细胞内CXCR3-mRNA和CXCR3蛋白表达水平;显著延长大鼠肝脏移植物存活期;asPNA组大鼠肝移植术后第7天外周血和肝脏细胞内CXCR3的表达较AR组及asPNA对照组明显减少（P〈0.05）,与FK506组呈现出类似的趋势。结论asPNA能有效抑制T细胞内CXCR3基因和蛋白表达水平,降低大鼠AR发生率并延长肝脏移植物存活期,具有潜在临床应用价值。     

全身照射预处理供体对异基因大鼠肝移植的作用及对受体内CD4~+CD25~+调节性T细胞的影响

目的 探讨全身照射(TBI)预处理诱导大鼠肝移植术后急性排斥反应的发生机制,及CD4~+ CD25~+调节性T细胞的变化在诱导免疫耐受中的作用.方法 以雄性Lewis、DA大鼠为供、受体,随机分为正常对照组、同种肝移植组、自发免疫耐受组、急性排斥反应组.观察各组受体的生存时间及生存率,检测受体术后外周血中ALT、TB含量、Foxp3~+ CD4~+ CD25~+ 调节性T细胞和T细胞亚群上GITR的表达,检测受体术后第14天移植肝的病理变化和受体脾脏CTL杀伤活性.结果 自发免疫耐受组,术后经历短暂排斥反应最终获得免疫耐受并长期存活.急性排斥反应组,在术后第17～21天死亡,与其他组相比,外周血血清中ALT、TB含量明显升高,而Foxp3~+ CD4~+ CD25~+调节性T细胞比例明显降低.TBI预处理大鼠供肝致受体外周血中CD3~+ CD4~+ T细胞上GITR表达降低,CD3~+CD8~+T细胞上GITR表达增加,提高CTL的杀伤活性.结论 通过TBI清除供体大鼠肝移植物中携带的旁路淋巴细胞,致受体外周血中Foxp3~+ CD4~+ CD25~+调节性T细胞表达降低,而使CD3~+ CD8~+T细胞上GITR表达增加,共同诱导大鼠肝移植术后急性排斥反应发生和耐受障碍.     

感染相关性噬血细胞综合征患者干扰素γ、IP-10和CXCR3的表达及临床意义

目的 探讨细胞因子干扰素γ（IFNγ）、干扰素诱导蛋白10（IP-10）及受体CXCR3在感染相关性噬血细胞综合征（infection-associated hemophagocytic syndrome,IAHPS）患者中的表达及临床意义.方法 采用酶联免疫吸附试验（ELISA）检测43例IAHPS患者血清IFNγ和IP-10的表达,以流式细胞术检测外周血CD_4~＋和CD_8~＋T淋巴细胞表面CXCR3的表达,并以无HPS感染组（35例）和健康体检者（20名）作为对照.采用SPSS 13.0软件分析数据,组间比较采用独立样本t检验.结果 IAHPS组血清IFNγ和IP-10水平分别为（608±135）pmol/L和（939±141）pmol/L,明显高于无HPS感染组[（154±45）pmol/L、（385±119）pmol/L,t值分别为4.97和4.02,P值均〈0.05]和健康对照组[（56±18）pmol/L、（248±98）pmol/L,t值分别为5.27和4.77,P值均〈0.05].IAHPS组的CD4＋和CD8＋T淋巴细胞表面CXCR3的表达分别为（35±11）%和（23±8）%,明显高于无HPS感染组[（24±7）%、（16±7）%,t值分别为3.12和3.62,P值均〈0.05]和健康对照组[（20±6）%、（12±5）%,t值分别为4.46和4.93,P值均〈0.05].结论 IAHPS患者外周血IFNγ和IP-10表达明显升高,CD_4~＋和CD_8~＋T淋巴细胞上的受体CXCR3表达也增强,这些趋化因子的异常表达可能与HPS的发病机制相关.     

环孢菌素A抑制大鼠同种心脏移植急性排斥反应中趋化因子受体CCR5表达的研究

目的观察大鼠同种心脏移植急性排斥反应中趋化因子受体CCID的表达及环孢菌素A（CsA）的抑制作用。方法分两组建立同种大鼠颈部心脏移植模型：对照组（n=25）和新山地明组（CsA组，n=25），各5例观察移植心脏存活时间。于移植术后第1、5、7、14天分别取移植心组织各5例，逆转录-聚合酶链反应（RT—PCR）检测移植心组织内趋化因子受体CCR5mRNA不同时间点的表达水平，免疫组织化学方法检测移植心组织内趋化因子受体CCR5分子的表达差异。结果对照组移植心存活时间为（11．6±1．5）d，CsA组移植心存活时间为（22．4±5．1）d，两组问移植心存活时间差异有统计学意义（P〈0．01）。急性排斥组和CsA处理组大鼠移植心脏在术后第5、7、14天都可检测到趋化因子受体CCR5mRNA阳性表达，但CsA处理组趋化因子受体CCRSmRNA的表达明显弱于急性排斥组。同样急性排斥组和CsA处理组大鼠移植心脏在术后第5、7、14天都可检测到趋化因子受体CCR5分子的表达，CsA处理组趋化因子受体CCR5分子的表达相对较弱。结论趋化因子受体CCR5在早期移植免疫事件中具有重要的作用，CsA能部分抑制趋化因子受体CCR5的表达，并与移植物存活时间延长有一定的关系。     

穿孔素mRNA表达与大鼠肝移植急性排斥反应的关系

目的 探讨大鼠肝脏移植术后外周血和胆汁中穿孔素mRNA表达水平与急性排斥反应的关系,寻求一种早期诊断急性排斥反应的非侵袭性方法.方法 建立大鼠原位肝移植模型及胆道引流模型,实验分为4组: 空白对照组(n=20),行单纯胆汁引流;同基因移植组(n=30);异基因移植+环孢素A(CsA)组(n=30);异基因移植组(n=30),术后未行免疫抑制剂治疗.应用RT-PCR方法分别检测各组术后第1、3、5、7和10天外周血和胆汁中穿孔素mRNA的表达水平,同时行肝脏病理组织学观察.结果 空白对照组、同基因移植组外周血中无穿孔素mRNA表达;异基因移植组外周血中穿孔素mRNA 在术后第3天开始表达,而后逐渐升高,术后第5天明显增高,术后第7～10天维持在较高水平.而异基因移植+CsA组外周血中穿孔素mRNA的表达持续呈低水平,各时间点表达与异基因移植组比较差异有统计学意义(P＜0.05).外周血穿孔素mRNA表达水平与Banff组织学诊断分级变化规律一致.各组胆汁中穿孔素mRNA表达均为阴性.结论 检测外周血中穿孔素mRNA表达可作为监测急性排斥反应较敏感的无创性方法.     

急性排斥反应时移植肾组织中趋化因子及其受体的表达

目的 探讨急性排斥反应时移植肾组织中正常T淋巴细胞表达及调节因子(RANTES)、单核细胞趋化因子-1(MCP-1)、γ干扰素诱导的单核因子(CXCL9/Mig)、γ干扰素诱导蛋白10(CXCL10/IP-10)及其受体CCR5、CCR2、CXCR3的表达及意义.方法 以52例肾移植受者为对象,移植后常规进行移植肾穿刺,所取材料一部分进行组织学观察,另一部分采用荧光定量聚合酶链反应(PCR)检测各趋化因子及其受体的mRNA表达.结果 52例中,17例为临界改变,21例为排斥改变,其中I a级18例,I b级3例(以上38例为排斥组).另14例肾功能稳定,移植肾组织无明显病理变化(非排斥组).排斥组RANTES、CXCL10及CCR5的mRNA表达明显上调,其mRNA表达量分别是非排斥组的2.70倍(P<0.01)、3.76倍(P<0.01)和6.88倍(P<0.01);两组间CXCL9、MCP-1及CCR2 mRNA表达的差异均无统计学意义(P>0.05).结论 急性排斥反应时,移植肾组织中RANTES、CXCL10和CCR5表达升高,提示它们在肾移植急性排斥反应中可能起重要作用,它们的高表达可以作为移植肾急性排斥反应的标志之一.     

Lewis型至Brown Norway型大鼠肝移植急性排斥反应模型的建立

目的:建立Lewis(LEN)型到Brown Norway(BN)型大鼠肝移植急性排斥反应模型并观察其排斥反应特点.方法:采用近交系雄性LEW大鼠为供体,BN大鼠为受体,改良"二袖套"法建立模型36例,术后第3、7和10天分别随机解剖8只受体大鼠,检测血清肝功能生化指标,观察肝脏病理组织学变化,剩余受体大鼠观察生存时间.结果:受体大鼠术后第3、7和10天的血清总胆红素分别为(8.75±1.98)μmol/L、(21.12±2.03)μmol/L和(53.63±4.24)μmol/L,丙氨酸转氨酶(ALT)分别为(291.88±6.83)U/L、(462.33±16.98)U/L和(906.25±68.55)U/L,两者术后均逐渐升高;受体大鼠术后第3天光镜下汇管区见混合性炎细胞浸润和静脉内皮炎,伴部分肝实质变性,排斥活动指数(RAI)为4～5;术后第7天排斥反应加重,并出现小胆管增生,RAI为5～7;术后第10天排斥反应最严重,出现明显汇管区结构破坏,肝小叶消失,大量淋巴细胞和单核细胞浸润,静脉内皮炎明显,肝实质桥接坏死,RAI为7～9分;受体大鼠的临床表现及生存情况与病理变化一致.结论:LEW到BN大鼠组合有助于建立稳定可靠的肝移植急性排斥模型,且该模型的排斥反应特点与临床相似.     

阻断B7/CD28共刺激通路抑制大鼠肝移植急性排斥反应的研究

目的:建立大鼠原位肝移植(ROLT)急性排斥反应模型,观察细胞毒淋巴细胞抗原4免疫球蛋白(CTLA-4Ig)在大鼠肝移植术后对共刺激分子B7-1/B7-2的影响及其抗排斥反应的作用。方法:采用"二袖套管"法先行建立DA-Lewis大鼠组合肝移植急性排斥反应模型,随机分为对照组(A组)与实验组(B组)两组,于肝移植术后48h每只受体大鼠腹腔内一次性注射CTLA-4Ig 75μg,分别于术后3、5、7和10d采用RT-PCR检测B7-1和B7-2 mRNA在两组肝脏组织中的表达情况,并同时观察其肝功能变化。结果:1)B7-1和B7-2 mRNA在A组高水平表达,而在B组表达明显降低(P<0.01);2)B组动物术后未见明显排斥反应,血清ALT、TBIL和DBIL水平明显低于对照组(P<0.01)。结论:移植术后应用CTLA-4Ig可以降低肝组织中B7-1和B7-2的表达;动态检测B7分子的表达有助于观察肝移植排斥反应的进程。     

大鼠肝移植物中T-bet+/GATA-3+和ROR-γt+/FOXP3+细胞随排斥程度的动态变化及意义

目的 检测Th1、Th2、Th17和iTreg CD4+T细胞亚系特征性表达的核转录因子以探究它们在大鼠肝移植急性排斥中的动态变化及意义.方法 Kamada"二袖套"法建立大鼠肝移植急性排斥模型,按Banff标准对排斥程度进行分级;免疫组织化学方法检测Th1、Th2、Th17和iTreg特征性表达的核转录因子,对阳性细胞计数.结果 与对照组比较,轻度排斥组、中度排斥组和重度排斥组中T-bet+(对照组:19.3±5.1;轻度排斥组:63.7±9.7;中度排斥组:40.9±13.6;重度排斥组:32.3±3.3)细胞数和RORγt+(对照组:6.5±1.4;轻度排斥组:8.3±2.3;中度排斥组:26.8±3.2;重度排斥组:39.2±12.8)细胞数显著升高(P<0.01);轻度排斥组中GATA-3+(对照组:7.1±1.3;轻度排斥组:6.4±3.1;中度排斥组:23.2±9.1;重度排斥组:42.6±14.1)细胞数无明显升高(P>0.05),中度排斥组中明显升高(P>0.05),重度排斥组显著升高(P<0.01);轻度排斥组FOXP3+(对照组:9.5±2.4;轻度排斥组:13.5±4.8;中度排斥组:24.5±4.9;重度排斥组:39.3±11.7)细胞数无明显升高(P>0.05),中度、重度排斥组显著升高(P<0.01).结论 大鼠肝移植术后急性排斥的发生与T-bet+、GATA-3+、RORγt+和FOXP3+细胞相关;在排斥早期,T-bet+/GATA-3+平衡与排斥的发生相关性比RORγt+/FOXP3+平衡更强.     

Serum Fractalkine and Interferon-Gamma Inducible Protein-10 Concentrations Are Early Detection Markers for Acute Renal Allograft Rejection

Objective

The aims of this study were to determine if characterization of serum concentrations of interferon-gamma inducible protein-10 (IP-10), fractalkine, and their receptors (CXCR3 and CX3CR1) were predictive of acute allograft rejection in kidney transplant recipients.

Methods

Kidney transplant recipients (n = 52) were enrolled in this study and divide into either the acute rejection (AR, n = 15) or non–acute rejection (NAR, n = 35) groups. Serum samples from recipients were collected 1 day prior to transplantation and on days 1, 3, 5, 7, and 9 post-transplantation. The accuracy of chemokine concentrations for predicting acute rejection episodes was evaluated using receiver operator characteristic (ROC) curves.

Results

AR was diagnosed in 15 patients based on histologic changes to renal biopsies. AR patients had significantly higher serum fractalkine, CXCR1, IP-10, and CXCR3 levels compared to levels observed in the NAR group and healthy controls. Fractalkine and IP-10 had the largest area under the ROC curve at 0.86 (95% confidence interval: 0.77–0.96). Following steroid therapy, chemokine levels decreased, which may serve to predict the therapeutic response to steroid therapy.

Conclusion

Measuring serum levels of fractalkine, IP-10, and their receptors (especially the fractalkine/IP-10 combination) may serve as a noninvasive approach for the early diagnosis of renal allograft rejection.     

Surveillance of acute rejection in baboon renal transplantation by elevation of interferon-gamma inducible protein-10 and monokine induced by interferon-gamma in urine

BACKGROUND: CXCR3 binding chemokines play a key role in recruitment of inflammatory cells into an organ transplant. This study addresses the question of whether urinary excretion of these chemokines correlates with acute rejection in a baboon kidney transplantation model. METHODS: Seven outbred baboons underwent renal allotransplantation from major histocompatibility complex (MHC)-mismatched donors. The treatment of baboons consisted of anti-CD4 monoclonal antibody (mAb), anti-CD8 mAb, rapamycin, and mycophenolate mofetil (MMF). Urinary levels of interferon-gamma inducible protein-10 (IP-10) and monokine induced by interferon-gamma (Mig) were determined by ELISA. Renal biopsies were examined by immunohistochemical staining for CXCR3 and Mig. RESULTS: Urinary levels of IP-10 and Mig increased significantly in all of the five baboons at the time of acute rejection of renal transplant. The IP-10 and Mig levels did not rise in two nonrejecting baboons. In two baboons, urinary levels of IP-10 and Mig rose before the elevation of the serum creatinine. In renal biopsies, expression of Mig was detected in glomeruli, tubules, and infiltrating cells, and the expression was significantly elevated in biopsies with acute rejection (P<0.01). CXCR3 was constitutively expressed in tubular cells in biopsies derived from both normal grafts and grafts with acute rejection. Whereas the infiltrating cells were increased in the biopsies with acute rejection, the expression of CXCR3 was also significantly higher (P<0.01) in these infiltrating cells compared with those in the normal controls. CONCLUSIONS: This study shows an important correlation between urinary excretion of IP-10 and Mig and acute rejection in baboon kidney transplantation.     

CXCR3 and CCR5 positive T-cell recruitment in acute human renal allograft rejection

BACKGROUND: Experimental studies suggest that the infiltration of activated T cells into the allograft, the key event in the development of acute renal allograft rejection, depends on the expression of chemokines and their interaction with chemokine receptors expressed on T cells. METHODS: For a more detailed comprehension of the pathogenesis of T-cell recruitment in human acute rejection, the in situ expression of chemokines and chemokine receptors in allografts of 26 patients between day 3 and 9 after renal transplantation was examined in the present prospective study. RESULTS: Immunohistochemical staining showed a significantly increased number of CXCR3 (P<0.01) and CCR5 positive T cells (P<0.01) in the tubulointerstitium of patients with acute allograft rejection according to Banff grade Ia-IIb. Likewise the intrarenal RNA expression of the CXCR3 ligands IP-10 (5.2-fold) and I-TAC (7.2-fold) and the CCR5 ligand RANTES (5.7-fold), was significantly up-regulated in rejecting organs. In situ hybridization revealed that IP-10 but not I-TAC was predominantly expressed by infiltrating leukocytes in the tubulointerstitial area, co-localizing with CXCR3 positive T cells. To a lesser degree expression by tubular cells could be detected, providing a possible explanation for the increased urinary IP-10 excretion we found in patients with rejecting organs. CONCLUSIONS: These data from a prospective, biopsy-controlled study indicate that the local expression of IP-10 and RANTES leads to the directional movement of activated CXCR3 and CCR5 bearing T cells into the renal allograft and mediates acute rejection. Our data provide a rationale that blocking CXCR3 and CCR5 may offer a unique therapeutic approach to prevent episodes of acute rejection in the early phase after renal transplantation.     

大鼠肝移植排斥反应时γ干扰素及白细胞介素10的表达及意义

目的 探讨大鼠肝移植排斥反应时γ干扰素(IFN-γ)及白细胞介素10(IL-10)的表达及意义.方法 采用改良的Kamada"二袖套法"制备大鼠原位肝移植模型,同系移植组供、受者均为SD大鼠;同种异体移植组的供者为Wistar大鼠,受者为SD大鼠;另设假手术组.术后7 d处死动物,观察移植肝脏的组织学变化,检测血清IFN-γ和IL-10的含量,以及移植肝脏内IFN-γ和IL-10 mRNA的表达.结果 同种异体移植组移植肝脏有较多坏死肝细胞,汇管区及中央静脉周围可见以淋巴细胞为主的炎症细胞浸润,胆管上皮细胞可见胞浆空泡变性、核固缩或碎裂,整个肝小叶结构紊乱.同系移植组肝脏组织结构仅有轻度缺血再灌注损伤表现,汇管区有较少炎症细胞浸润,胆管上皮细胞结构和肝小叶结构基本正常.同种异体移植组血清IFN-γ为(386.7±14.4)Pg/ml,明显高于同系移植组的(159.8±16.5)pg/ml(P<0.05);同种异体移植组血清IL-10为(126.3±13.1)pg/ml,明显低于同系移植组的(288.3±17.1)pg/ml(P<0.05).同种异体移植组移植肝组织内IF-γ mRNA表达水平明显高于同系移植组(P<0.05),而IL-10 rnRNA表达水平明显低于同系移植组(P<0.05).结论 大鼠肝移植排斥反应时IFN-γ表达明显升高,IL-10表达明显降低;T_H1/T_H2型细胞因子的动态平衡可能在大鼠肝移植排斥反应中起着重要作用.     

IFN-inducible protein-10 has a differential role in podocyte during Thy 1.1 glomerulonephritis

IFN-inducible protein-10 (IP-10/CXCL10) is a potent chemoattractant for activated T lymphocytes and was recently reported to have several additional biologic activities. In this study, the expression and the function in normal glomeruli and in Thy1.1 glomerulonephritis (GN) were investigated. The expression of IP-10 was detected in normal rat glomeruli mainly in the podocyte. The expression of IP-10 was also detected on the cultured podocyte. The IP-10 expression was elevated at the early phase of Thy1.1 GN. The double staining immunofluorescence study clearly demonstrated that the elevated expression of IP-10 was mostly detected in the podocyte and very partly in mesangial area. A receptor for IP-10, CXCR3, showed similar expression patterns to that of IP-10. Expressions of neither of IP-10 nor of CXCR3 were detected on the inflammatory cells. For elucidating the role of IP-10, the blocking study was carried out with monoclonal anti-IP-10 antibody. The monoclonal anti-IP-10 antibody treatment decreased the expression of IP-10 and podocyte-associated proteins such as nephrin and podocin that are reported to be essential for maintaining the podocyte function (IP-10, 53.0% to control; nephrin, 43.5%; podocin, 60.4%). The findings indicated that the anti-IP-10 treatment disturbed the podocyte function. The anti-IP-10 treatment given to the rats with Thy1.1 nephritis exacerbated proteinuria, mesangiolysis, and matrix expansion. Collectively, the findings indicated that IP-10 plays a role in maintaining the podocyte function. Also, the findings suggested that anti-IP-10 treatment exacerbated the glomerular alterations in Thy1.1 GN by disturbing the podocyte function.     

FK330, a Novel Inducible Nitric Oxide Synthase Inhibitor, Prevents Ischemia and Reperfusion Injury in Rat Liver Transplantation

Nitric oxide (NO), produced via inducible NO synthase (iNOS), is implicated in the pathophysiology of liver ischemia/reperfusion injury (IRI). We examined the effects of a novel iNOS inhibitor, FK330 (FR260330), in well-defined rat liver IRI models. In a model of liver cold ischemia followed by ex vivo reperfusion, treatment with FK330 improved portal venous flow, increased bile production and decreased hepatocellular damage. FK330 prevented IRI in rat model of 40-h cold ischemia followed by syngeneic orthotopic liver transplantation (OLT), as evidenced by: (1) increased OLT survival (from 20% to 80%); (2) decreased hepatocellular damage (serum glutamic oxaloacetic transaminase/glutamic pyruvic transaminase levels); (3) improved histological features of IRI; (4) reduced intrahepatic leukocyte infiltration, as evidenced by decreased expression of P-selectin/intracellular adhesion molecule 1, ED-1/CD3 cells and neutrophils; (5) depressed lymphocyte activation, as evidenced by expression of pro-inflammatory cytokine (TNF-alpha, IL-1beta, IL-6) and chemokine (IP-10, MCP-1, MIP-2) programs; (6) prevented hepatic apoptosis and down-regulated Bax/Bcl-2 ratio. Thus, by modulating leukocyte trafficking and cell activation patterns, treatment of rats with FK330, a specific iNOS inhibitor, prevented liver IRI. These results provide the rationale for novel therapeutic approaches to maximize organ donor pool through the safer use of liver grafts despite prolonged periods of cold ischemia.     

Selective neutralization of the chemokine TCA3 reduces the increased injury of partial versus whole liver transplants induced by cold preservation

BACKGROUND: Given the shortage of liver donors and the development of techniques for partial liver transplantation, we compared chemokine expression and inflammatory cell infiltration of partial versus whole grafts in a mouse syngeneic liver transplant model. METHODS: Orthotopic liver transplantation, using whole or partial murine liver grafts, was performed following cold preservation in ViaSpan solution for periods of one to eight hours. RESULTS: Partial grafts showed more severe cold ischemia/reperfusion injury and greater inflammatory cell infiltration than whole grafts, and was accompanied by the marked intrahepatic upregulation of multiple chemokines. Quantitative analysis showed that compared with expression in whole grafts harvested after the same period of cold ischemia, partial grafts had eightfold more T-cell activation gene (TCA)-3 (CCL1) chemokine messenger RNA (mRNA) expression (P<0.01) and sixfold more inducible protein (IP)-10 chemokine (CCL10) mRNA expression (P<0.01), as well as increased expression of the chemokine receptors CCR8 (receptor for TCA3) and CXCR3 (receptor for IP-10; P<0.01). Blockade of TCA3 by neutralizing monoclonal antibody significantly decreased intragraft IP-10 expression (P<0.05) but not tumor necrosis factor-alpha or interleukin-6 expression in partial grafts, and significantly decreased cold ischemia/reperfusion injury (P<0.05) and associated neutrophil and T-cell infiltration (P<0.01). CONCLUSIONS: These data demonstrate that the chemokine TCA3/CCL1 is important to the pathogenesis of ischemic injury of experimental partial liver grafts, and that its therapeutic targeting within such grafts can overcome the deleterious effects of prolonged cold preservation and restore liver function to the level achieved using whole liver grafts.     

褪黑素对大鼠移植肝组织GRP-78及CHOP表达的影响

目的 通过检测褪黑素(MEL)对肝移植大鼠肝脏内质网应激(ERS)通路相关分子葡萄糖调节蛋白-78(GRP-78)及CCAAT增强子结合蛋白同源蛋白(CHOP)表达的影响, 探讨MEL对移植肝的作用。 方法 采用"磁环法"制作大鼠原位肝移植模型, 雄性SD大鼠随机分为3组:假手术组(Sham组)、原位肝移植组(OLT组)、原位肝移植+褪黑素处理组(OLT+MEL组), 每组各8只。术后24 h, 获取血清样本及肝脏标本。检测血清丙氨酸转移酶(ALT)及天冬氨酸转移酶(AST)水平, 苏木素-伊红(HE)染色观察各组肝组织病理学改变, 采用聚合酶链反应和蛋白免疫印迹试验检测GRP-78及CHOP的信使核糖核酸(mRNA)和蛋白表达水平。 结果 与Sham组比较, OLT组血清ALT、AST显著升高(均为P < 0.01), 肝组织损伤严重, GRP-78及CHOP在mRNA和蛋白水平表达明显增加(均为P < 0.01)。与OLT组比较, OLT+MEL组大鼠ALT、AST水平明显降低(均为P < 0.05), 肝组织损伤减轻, GRP-78及CHOP mRNA和蛋白表达水平明显降低(均为P < 0.05)。 结论 褪黑素能降低移植肝ERS相关分子GRP-78及CHOP在mRNA和蛋白水平的表达, 这可能是它减轻移植后肝脏损伤的机制之一。