α-ketoglutarate attenuates ischemia-reperfusion injury of liver graft in rats
α-ketoglutarate attenuates ischemia-reperfusion injury of liver graft in rats
复制标题
α-酮戊二酸减轻大鼠肝移植缺血再灌注损伤
DOI：
10.1016/j.biopha.2018.12.149
复制
发表时间：
2019-03-01
影响因子：
7.5
通讯作者：
Gong, Jian-ping
中科院分区：
文献类型：
--
作者：
Cheng, Ming-xiang;Cao, Ding;Gong, Jian-ping
目的：α-酮戊二酸（alpha-ketoglutarate，alphaKG）是巨噬细胞活化的重要代谢产物。本研究旨在探讨α KG /γ-氨基乙醇代谢是否能抑制肝移植过程中枯否细胞（KCs）的活化，减轻肝脏缺血再灌注损伤（IRI）。方法：供肝经门静脉灌注α KG的细胞渗透性类似物DM-α KG或γ-氨基乙醇酶1抑制剂BPTES，对照组灌注UW液。建立大鼠肝移植模型。24 h后分析血清丙氨酸转氨酶（ALT）、总胆红素（TBIL）和炎性细胞因子水平以及组织学。从移植物中分离KC。RT-PCR和免疫荧光法检测极化特异性标记基因。Western blot检测糖原合成酶激酶3 β（p-GSK 3 β）磷酸化和细胞因子信号转导抑制因子1（SOCS 1）的表达。结果：与对照组相比，DM-α B灌流可降低ALT和TBIL水平，减轻肝损伤，减少细胞凋亡，而BPTES组ALT和TBIL水平较高，损伤严重，细胞凋亡较多。此外，DM-α KG灌注抑制NF-κ B活性，上调KC中p-GSK 3 β和SOCS 1的表达，并使M1/M2平衡向抗炎性方向转变。DM-alpha KG可抑制血清促炎细胞因子的分泌，增加IL-10的表达。结论：α KG通过调节炎症反应和改变KCs的极化来保护移植肝免受IRI的影响。
Objective: The alpha-ketoglutarate (alpha KG), a metabolite of glutaminolysis, is reported to orchestrate macrophages activation. This study aims to clarify whether the alpha KG / glutaminolysis metabolism can suppress Kupffer cells (KCs) activation during liver transplantation and attenuate hepatic ischemia-reperfusion injury (IRI).Methods: Donor livers were perfused with DM-alpha KG (a cell-permeable analog of alpha KG) or BPTES (an inhibitor of glutaminase 1) via portal vein during cold preservation, and controls were perfused with UW solution. Then, a rat model of liver transplantation was performed. Serum levels of alanine transaminase (ALT), total bilirubin (TBIL) and inflammatory cytokines, as well as histology, were analyzed after 24 h. KCs were isolated from grafts. RT-PCR and immunofluorescence were used to evaluate polarization-specific marker genes. Western bolt was employed to assess the expression of phosphorylation of glycogen synthase kinase 3 beta (p-GSK3 beta) and suppressor of cytokine signaling 1 (SOCS1). EMSA was utilized to quantify the NF-kappa B transcriptional activity.Results: Compared with controls, DM-alpha KG perfusion decreased ALT and TBIL levels, alleviated liver damage, and reduced apoptosis, while BPTES group showed higher ALT and TBIL levels, severe damage and more apoptosis. Furthermore, DM-alpha KG perfusion suppressed NF-kappa B activity, up-regulated the expression of p-GSK3 beta and SOCS1 in KCs, and shifted the M1/M2 balance toward an anti-inflammatory profile. Besides, DM-alpha KG suppressed serum pro-inflammatory cytokines secretion and increased IL-10.Conclusions: alpha KG produced by glutaminolysis protects liver graft from IRI by regulating the inflammatory response and modifying the polarization of KCs.