Native Macrophages Genetically Modified to Express Heme Oxygenase 1 Protect Rat Liver Transplants from Ischemia/Reperfusion Injury
Native Macrophages Genetically Modified to Express Heme Oxygenase 1 Protect Rat Liver Transplants from Ischemia/Reperfusion Injury
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DOI：
10.1002/lt.22214
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发表时间：
2011-02-01
影响因子：
4.6
通讯作者：
Kupiec-Weglinski, Jerzy W.
中科院分区：
文献类型：
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作者：
Shen, Xiu-Da;Ke, Bibo;Kupiec-Weglinski, Jerzy W.
我们研究了天然巨噬细胞过度表达血红素氧合酶1（HO-1）是否可以保护大鼠原位肝移植（奥尔特）免受冷缺血/再灌注损伤（IRI）。来自Sprague-Dawley大鼠的肝脏在威斯康星州大学的溶液中于4 ℃下储存24小时，然后将它们移植到同基因受体中。在再灌注前将用血红素加氧酶1腺病毒（Ad-HO-1）、β-半乳糖苷酶腺病毒（Ad-beta-gal）或HO-1小干扰RNA（siRNA）离体转染的骨髓源性巨噬细胞（BMSCs）直接注入奥尔特。对照是用未修饰的或乱序siRNA转染的细胞调节的奥尔特。Ad-HO-1/Bcl-2的转移使奥尔特的存活率提高至100%（对照组为40%-50%），并降低血清丙氨酸氨基转移酶水平和肝细胞损伤的组织学特征。相反，输注HO-1 siRNA/Ad-beta-gal转染的巨噬细胞未能影响IRI。基因治疗诱导HO-1抑制Toll样受体4的表达，降低促炎性肿瘤坏死因子α、白细胞介素-1 β、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，并减弱内皮细胞间粘附分子1的表达，从而减少奥尔特白细胞隔离。尽管Ad-HO-1/Bcl 4可降低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凋亡细胞的频率，并改善caspase-3活性，但在功能良好的奥尔特中，白细胞介素-10和抗凋亡B细胞淋巴瘤2/B细胞淋巴瘤特大型的表达增加。因此，用HO-1离体转染的天然巨噬细胞的转移可以从IRI中拯救大鼠iso-OLT。我们的研究验证了一个新的和临床上有吸引力的概念：与抗氧化剂HO-1体外转染的天然巨噬细胞可以在移植时应用，以减轻否则破坏性的抗原非依赖性肝脏炎症和损伤所造成的周围transplant收获侮辱。如果这种新的，完善的策略被证明是有效的allo-OLT受体，它应该被考虑在临床环境中，以增加可用的供体器官的供应，并最终提高肝移植的整体成功率。肝移植17：201-210，2011. (C)2011年AASLD。
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 degrees 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), beta-galactosidase adenovirus (Ad-beta-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-beta-gal failed to affect IRI. Gene therapy induced HO-1 suppressed toll-like receptor 4 expression, decreased expression of proinflammatory tumor necrosis factor alpha, interleukin-1 beta, 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. Liver Transpl 17:201-210, 2011. (C) 2011 AASLD.