Ke Bibo, Shen Xiu-Da, Tsuchihashi Sei-Ichiro, Gao Feng, Araujo Jesus A, Busuttil Ronald W, Ritter Thomas, Kupiec-Weglinski Jerzy W
Dumont-UCLA Transplant Center, Department of Surgery, David Geffen School of Medicine at UCLA, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Hum Gene Ther. 2007 Apr;18(4):355-66. doi: 10.1089/hum.2007.181.
Ischemia-reperfusion injury (IRI) contributes to early and late dysfunction of liver transplants. We have shown that sentinel Toll-like receptor-4 (TLR4) plays a key role in the activation of T cell immune responses during hepatic IRI. We have also documented that overexpression of heme oxygenase-1 (HO-1) exerts potent cytoprotective effects. This study analyzes how adenovirus (Ad)-based viral interleukin-10 (vIL-10) gene transfer affects TLR4 and HO-1 signaling in host innate and adaptive immunity during liver IRI. Using a partial lobar warm IRI model, groups of wild-type and HO-1(+/-) knockout (KO) mice were assessed for severity of hepatocellular damage after 90 min of warm ischemia followed by 6 hr of reperfusion. Both wild-type and HO-1 (+/-) KO mice treated with Ad-vIL-10 have shown improved hepatic function (serum glutamic-oxaloacetic transaminase levels), ameliorated histological signs of IRI (Suzuki's score), decreased neutrophil accumulation (myeloperoxidase activity), and depressed tumor necrosis factor-alpha/IL-1beta, IL-2/interferon-gamma, E-selectin, and macrophage inflammatory protein-2 expression. These effects were IL-10 dependent as treatment with neutralizing antibody re-created liver IRI. In contrast, untreated wild-type and HO-1 (+/-) KO mice, as well as wild-type and HO-1 (+/-) KO mice treated with Ad-beta-Gal, showed severe hepatocellular damage due to IRI. Unlike in controls, wild-type and HO-1 (+/-) KO mice treated with Ad-vIL-10 revealed markedly depressed TLR4 and NF-kappaB expression, along with increased HO-1 and Bcl-2/Bcl-x(L) expression, as compared with respective controls. Thus, vIL-10 gene transfer prevents hepatic IRI in association with depressed expression of innate TLR4, and adaptive Th1 cytokine/chemokine programs. The induction of antioxidant HO-1 and anti-apoptotic Bcl-2/Bcl-x(L) by vIL-10 exerts synergistic cytoprotective function against antigen-independent hepatic inflammatory response triggered by IRI.
缺血再灌注损伤（IRI）会导致肝移植的早期和晚期功能障碍。我们已经表明，前哨Toll样受体4（TLR4）在肝脏IRI期间T细胞免疫反应的激活中起关键作用。我们还记录到，血红素加氧酶-1（HO-1）的过表达具有强大的细胞保护作用。本研究分析了基于腺病毒（Ad）的病毒白细胞介素-10（vIL-10）基因转移如何影响肝脏IRI期间宿主固有免疫和适应性免疫中的TLR4和HO-1信号传导。使用部分肝叶热缺血再灌注损伤模型，对野生型和HO-1(+/-)基因敲除（KO）小鼠组在进行90分钟热缺血后再灌注6小时后评估肝细胞损伤的严重程度。用Ad-vIL-10处理的野生型和HO-1(+/-) KO小鼠均显示肝功能改善（血清谷氨酸草酰乙酸转氨酶水平）、IRI的组织学迹象改善（铃木评分）、中性粒细胞积聚减少（髓过氧化物酶活性）以及肿瘤坏死因子-α/白细胞介素-1β、白细胞介素-2/干扰素-γ、E-选择素和巨噬细胞炎性蛋白-2表达降低。这些作用是IL-10依赖性的，因为用中和抗体处理会再次引发肝脏IRI。相比之下，未处理的野生型和HO-1(+/-) KO小鼠以及用Ad-β-半乳糖苷酶处理的野生型和HO-1(+/-) KO小鼠由于IRI而表现出严重的肝细胞损伤。与对照组不同，用Ad-vIL-10处理的野生型和HO-1(+/-) KO小鼠与各自的对照组相比，显示出TLR4和核因子-κB表达明显降低，同时HO-1和Bcl-2/Bcl-x(L)表达增加。因此，vIL-10基因转移可预防肝脏IRI，这与固有TLR4的表达降低以及适应性Th1细胞因子/趋化因子程序有关。vIL-10诱导抗氧化剂HO-1和抗凋亡蛋白Bcl-2/Bcl-x(L)对IRI引发的抗原非依赖性肝脏炎症反应发挥协同细胞保护作用。
