Department of Surgery, David Geffen School of Medicine, University of California Los Angeles, Dumont-UCLA Transplant Center, Los Angeles, CA 90095, USA.
Liver Transpl. 2011 Feb;17(2):201-10. doi: 10.1002/lt.22214.
We investigated whether native macrophages overexpressing heme oxygenase 1 (HO-1) could protect rat orthotopic liver transplant (OLT) against cold ischemia/reperfusion injury (IRI). Livers from Sprague-Dawley rats were stored at 4°C in University of Wisconsin solution for 24 hours, and then they were transplanted into syngeneic recipients. Bone marrow-derived macrophages (BMMs) that were transfected ex vivo with heme oxygenase 1 adenovirus (Ad-HO-1), β-galactosidase adenovirus (Ad-β-gal), or HO-1 small interfering RNA (siRNA) were infused directly into the OLT before reperfusion. Controls were OLT conditioned with unmodified or scrambled siRNA-transfected cells. The transfer of Ad-HO-1/BMMs increased the survival of OLT to 100% (versus 40%-50% for controls) and decreased serum alanine aminotransferase levels and histological features of hepatocellular damage. In contrast, an infusion of macrophages transfected with HO-1 siRNA/Ad-β-gal failed to affect IRI. Gene therapy-induced HO-1 suppressed toll-like receptor 4 expression, decreased expression of proinflammatory tumor necrosis factor α, interleukin-1β, monocyte chemoattractant protein 1, and chemokine (C-X-C motif) ligand 10, and attenuated endothelial intercellular cell adhesion molecule 1 expression with resultant diminished OLT leukocyte sequestration. Although Ad-HO-1/BMMs decreased the frequency of apoptotic cells positive for terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and ameliorated caspase-3 activity, the expression of interleukin-10 and antiapoptotic B cell lymphoma 2/B cell lymphoma extra large increased in well-functioning OLT. Thus, the transfer of native macrophages transfected ex vivo with HO-1 can rescue rat iso-OLT from IRI. Our study validates a novel and clinically attractive concept: native macrophages transfected ex vivo with the antioxidant HO-1 can be applied at the time of transplantation to mitigate otherwise damaging antigen-independent liver inflammation and injury resulting from the peritransplant harvesting insult. If this new, refined strategy is proven to be effective in allo-OLT recipients, it should be considered in clinical settings to increase the supply of usable donor organs and ultimately improve the overall success of liver transplantation.
我们研究了过表达血红素加氧酶 1（HO-1）的天然巨噬细胞是否可以防止大鼠原位肝移植（OLT）免受冷缺血/再灌注损伤（IRI）。Sprague-Dawley 大鼠的肝脏在 4°C 的威斯康星大学溶液中保存 24 小时，然后移植到同基因受体中。骨髓来源的巨噬细胞（BMMs）在体外转染血红素加氧酶 1 腺病毒（Ad-HO-1）、β-半乳糖苷酶腺病毒（Ad-β-gal）或 HO-1 小干扰 RNA（siRNA）后，在再灌注前直接输注到 OLT 中。对照组为用未修饰或乱序 siRNA 转染细胞预处理的 OLT。Ad-HO-1/BMM 的转移将 OLT 的存活率提高到 100%（对照组为 40%-50%），并降低了血清丙氨酸氨基转移酶水平和肝细胞损伤的组织学特征。相比之下，输注转染 HO-1 siRNA/Ad-β-gal 的巨噬细胞对 IRI 没有影响。基因治疗诱导的 HO-1 抑制 Toll 样受体 4 的表达，降低促炎肿瘤坏死因子 α、白细胞介素-1β、单核细胞趋化蛋白 1 和趋化因子（C-X-C 基序）配体 10 的表达，并减弱内皮细胞间细胞黏附分子 1 的表达，从而减少 OLT 白细胞的滞留。尽管 Ad-HO-1/BMM 减少了末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记阳性的凋亡细胞的频率，并改善了半胱氨酸天冬氨酸蛋白酶 3 的活性，但在功能良好的 OLT 中，白细胞介素-10 和抗凋亡 B 细胞淋巴瘤 2/B 细胞淋巴瘤 extra large 的表达增加。因此，体外转染 HO-1 的天然巨噬细胞的转移可以挽救大鼠同种异体 OLT 免受 IRI。我们的研究验证了一个新的、有吸引力的临床概念：体外转染抗氧化剂 HO-1 的天然巨噬细胞可以在移植时应用，以减轻移植采集损伤引起的、否则会造成损害的抗原非依赖性肝炎症和损伤。如果这种新的、精细的策略被证明在同种异体 OLT 受体中有效，那么它应该在临床环境中得到考虑，以增加可用供体器官的供应，最终提高肝移植的整体成功率。
