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 machine perfusion,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(alanine aminotransferase,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。
Objective This study aims to observe the reparative effect of Necrostatin-1(NEC-1)on a 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 the perfusion 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 higher than 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.
作者
孙平
吕佳蕊
王希
程颖
Sun Ping;Lv Jiarui;Wang Xi;Cheng Ying(Organ Transplantation and Hepatobiliary Surgery Department,First Affiliated Hospital of China Medical University,Liaoning Shenyang 110000,China)
出处
《实用器官移植电子杂志》
2025年第4期325-330,共6页
Practical Journal of Organ Transplantation(Electronic Version)
基金
国家重点研发计划课题任务书(2024YFA1109002)
辽宁省中央引导地方科技发展专项项目任务合同书(2024JH6/100800024)。
