Department of Surgery, University of Alberta, Edmonton, Canada.
Department of Pathology, University of Alberta, Edmonton, Canada.
PLoS One. 2019 Nov 26;14(11):e0224567. doi: 10.1371/journal.pone.0224567. eCollection 2019.
Liver ischemia reperfusion injury (IRI) remains a challenge in liver transplantation. A number of compounds have previously demonstrated efficacy in mitigating IRI. Herein, we applied three specific additive strategies to a mouse IRI screening model to determine their relative potencies in reducing such injury, with a view to future testing in a large animal and clinical ex situ normothermic perfusion setting: 1) F573, a pan-caspase inhibitor, 2) anti-inflammatory anakinra and etanrecept and 3) BMX-001, a mimetic of superoxide dismutase.
A non-lethal liver ischemia model in mice was used. Additives in the treatment groups were given at fixed time points before induction of injury, compared to a vehicle group that received no therapeutic treatment. Mice were recovered for 6 hours following the ischemic insult, at which point blood and tissue samples were obtained. Plasma was processed for transaminase levels. Whole liver tissue samples were processed for histology, markers of apoptosis, oxidative stress, and cytokine levels.
In an in vivo murine IRI model, the F573 treatment group demonstrated statistically lower alanine aminotransferase (ALT) levels (p = 0.01), less evidence of apoptosis (p = 0.03), and lower cytokine levels compared to vehicle. The etanercept with anakinra treatment group demonstrated significantly lower cytokine levels. The BMX-001 group demonstrated significantly decreased apoptosis (p = 0.01) evident on TUNEL staining.
The administration of pan-caspase inhibitor F573 in a murine in vivo model likely mitigates liver IRI based on decreased markers of cellular injury, decreased evidence of apoptosis, and improved cytokine profiles. Anakinra with etanercept, and BMX-001 did not demonstrate convincing efficacy at reducing IRI in this model, and likely need further optimization. The positive findings set rational groundwork for future translational studies of applying F573 during normothermic ex situ liver perfusion, with the aim of improving the quality of marginal grafts.
肝缺血再灌注损伤（IRI）仍然是肝移植中的一个挑战。以前有许多化合物在减轻 IRI 方面显示出了疗效。在此，我们将三种特定的添加策略应用于小鼠 IRI 筛选模型，以确定它们在减少这种损伤方面的相对效力，以期在大型动物和临床离体常温灌流环境中进行进一步的测试：1）F573，一种泛半胱天冬酶抑制剂，2）抗炎药阿那白滞素和依那西普，3）BMX-001，一种超氧化物歧化酶模拟物。
采用非致死性小鼠肝缺血模型。在诱导损伤前的固定时间点给予治疗组中的添加物，与未接受任何治疗的载体组进行比较。在缺血损伤后 6 小时恢复小鼠，此时获得血液和组织样本。处理血浆以测定转氨酶水平。处理整个肝组织样本以进行组织学、细胞凋亡、氧化应激和细胞因子水平的检测。
在体内小鼠 IRI 模型中，F573 治疗组的丙氨酸氨基转移酶（ALT）水平显著降低（p=0.01），细胞凋亡的证据更少（p=0.03），细胞因子水平也更低。依那西普与阿那白滞素联合治疗组的细胞因子水平显著降低。BMX-001 组的 TUNEL 染色显示凋亡明显减少（p=0.01）。
在体内小鼠模型中，泛半胱天冬酶抑制剂 F573 的给药可能通过降低细胞损伤标志物、减少细胞凋亡证据和改善细胞因子谱来减轻肝 IRI。阿那白滞素与依那西普联合治疗组和 BMX-001 组在该模型中并未显示出令人信服的减轻 IRI 的疗效，可能需要进一步优化。这些积极的发现为今后在常温离体肝脏灌流中应用 F573 进行转化研究奠定了合理的基础，旨在改善边缘供体的质量。
