Abstract:

Objective This study aims to observe the reparative effect of Necrostatin-1（NEC-1）ona rat model of moderate steatotic donor liver during NMP，and to preliminarily explore the mechanism of drug repair，providing experimental evidence for optimizing NMP strategies for steatotic donor livers. Methods This experiment included a total of 33 SD rats（male，8 ～ 10 weeks old），with 11 randomly selected for obtaining nonsteatotic liver as the liver source of animals，and the remaining 22 fed a high-fat diet for 2 months to induce steatotic liver confirmed by frozen section Oil Red O staining. The rats were divided into 3 groups ：Lean liver group（L group， n ＝ 6），steatotic liver group（S group，n ＝ 6），and steatotic liver group with addition of Necrostatin-1 to theperfusion solution（NS group，n ＝ 6），the 3 groups were preserved on a normothermic machine perfusion platform for 4 hours and analyzed by the infusion solution and liver tissue. Results During the perfusion process，the enzymatic elevation changes of NS group and S group were significantly higher than those of L group（P ＜ 0.05），NEC-1 had no significant effect on enzymatic changes（P ＞ 0.05），and the lactate concentration of NS group and S group was significantly higher than that of L group（P ＜ 0.05），but the lactate metabolism of NS group was effectively improved（P ＜ 0.05）. The triglyceride output of the NS group and the S group was significantly higherthan that of the L group（P ＜ 0.05），and the effect of NEC-1 on the secretion of triglycerides was not significant （P ＞ 0.05）. Conclusion Necrostatin-1 was able to improve the metabolic level of the rat model of steatosis donor liver in the experimental model of NMP ，and the repair effect of the drug on steatosis donor liver may be related to the downregulation of the necroptotic pathway. 

Key words:  , Steatosis donor liver, Necrostatin-1, Normothermic machine perfusion, Necroptosis

摘要：

目的 本实验旨在观察应用 Necrostatin-1（NEC-1）在常温机械灌注（normothermic machineperfusion，NMP）模型中对大鼠中度脂肪变性肝脏的修复作用，并初步探索药物修复的机制，为优化脂肪变性供肝常温机械灌注方案提供一定的实验依据。 方法 本研究选取 33 只 8 ～ 10 周龄的雄性 SD 大鼠，其中 11 只作为非脂肪变性供肝，22 只经高脂饲料喂养 2 个月后确认中度脂肪变性。大鼠分为三组 ：非脂肪变性供肝（L 组）、脂肪变性供肝（S 组）和添加 NEC-1 的脂肪变性供肝（NS 组）。各组在常温机械灌注下保存 4 h，并收集灌注液及肝组织进行分析。研究内容为肝组织 HE 染色、丙氨酸转氨酶（alanineaminotransferase，ALT）/ 天冬氨酸转氨酶（aspartate aminotransferase，AST）、乳酸和甘油三酯浓度变化，以及坏死性凋亡信号的比较。 结果 三组供肝在灌注过程中，NS 组和 S 组酶学升高变化显著高于 L 组（P ＜ 0.05），NEC-1 对酶学变化无显著影响（P ＞ 0.05）。NS 组和 S 组乳酸浓度显著高于 L 组（P ＜ 0.05），但 NS 组乳酸代谢有效改善（P ＜ 0.05）。NS 组和 S 组甘油三酯输出显著高于 L 组（P ＜ 0.05），NEC-1 对甘油三酯分泌影响不显著（P ＞ 0.05）。肝组织病理显示 NS 组和 S 组较 L 组脂肪变性严重，NS 组炎性浸润轻于 S 组。肝组织蛋白切迹显示 NS 组和 S 组较 L 组坏死性凋亡分子表达增强，但 NEC-1 能够降低坏死性凋亡分子表达。 结论 在本大鼠离体供肝 NMP 的实验模型中，Necrostatin-1 能够改善大鼠中度脂肪变性模型的代谢水平，该药物对脂肪变性供肝的修复作用可能与坏死性凋亡通路的下调有关。 

关键词:

脂肪变性供肝 , Necrostatin-1 , 常温机械灌注平台 , 坏死性凋亡 ,