Methyl eugenol attenuates liver ischemia reperfusion injury via activating PI3K/Akt signaling
Methyl eugenol attenuates liver ischemia reperfusion injury via activating PI3K/Akt signaling
复制标题
甲基丁香酚通过激活 PI3K/Akt 信号传导减轻肝脏缺血再灌注损伤
DOI：
10.1016/j.intimp.2021.108023
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发表时间：
2021-08-03
影响因子：
5.6
通讯作者：
Gong, Nianqiao
中科院分区：
文献类型：
--
作者：
Wang, Mengqin;Zhang, Ji;Gong, Nianqiao
背景资料：肝缺血再灌注损伤（liver ischemia reperfusion injury，LIRI）是肝移植、肝切除和各种循环休克过程中常见的损伤，可导致严重的代谢紊乱、炎性免疫反应、氧化应激损伤和细胞凋亡。甲基丁香酚（ME）在结构上与丁香酚相似，并具有抗肿瘤和凋亡的药理作用。然而，ME是否保护肝脏免受LIRI损伤还需要进一步研究。方法：采用C57BL/6J小鼠缺血60 min，再灌注6 h，建立部分温热LIRI模型。我们还建立了缺氧-复氧损伤（H/R）细胞模型，使AML 12（小鼠肝细胞系）细胞缺氧24 h，然后18 h常氧。通过血清转氨酶浓度、苏木精和伊红染色、实时定量PCR、髓过氧化物酶活性和TUNEL分析评估肝损伤程度。流式细胞术检测细胞凋亡。Western blotting检测p-PI3K、PI3K、p-Akt、Akt、p-Bad、Bad、Bcl-2、Bax和切割型caspase-3蛋白水平。用PI3K/Akt信号通路抑制剂LY294002研究ME与PI3K/Akt信号通路的关系。结果如下：ME能明显减轻LIRI诱导的肝损伤、炎症反应、细胞凋亡及缺氧/复氧诱导的肝细胞损伤。已知ME在体内和体外肝细胞损伤中激活PI3K/Akt信号通路，并且当该信号通路被抑制时，ME的保护作用被废除。结论：ME是一种通过激活PI3K/Akt信号调节LIRI的潜在治疗方法。
Background: Liver ischemia reperfusion injury (LIRI) often occurs during liver transplantation, resection, and various circulatory shock procedures, leading to severe metabolic disorders, inflammatory immune responses, oxidative stress injury, and cell apoptosis. Methyl eugenol (ME) is structurally similar to eugenol and has antiinflammatory and apoptotic pharmacological effects. However, whether ME protects the liver from LIRI damage requires further investigation. Methods: We established a partially warm LIRI model by subjecting C57BL/6J mice to 60 min of ischemia, followed by reperfusion for 6 h. We also established a hypoxia-reoxygenation injury (H/R) cell model by subjecting AML12 (a mouse liver cell line) cells to 24 h hypoxia, followed by 18 h normoxia. The extent of liver injury was assessed by serum transaminase concentrations, hematoxylin and eosin staining, quantitative real-time PCR, myeloperoxidase activity, and TUNEL analysis. Apoptosis was detected using flow cytometry. The protein levels of p-PI3K, PI3K, p-Akt, Akt, p-Bad, Bad, Bcl-2, Bax, and cleaved caspase-3 were detected by western blotting. LY294002, an inhibitor of PI3K/Akt signaling, was used to elucidate the relationship between ME and PI3K/Akt signaling. Results: ME successfully alleviated LIRI-induced liver injury, inflammatory response, and apoptosis induced, as well as liver cell injury induced by hypoxia reoxygenation. ME is known to activate the PI3K/Akt signaling pathway in hepatocyte injury in vivo and in vitro, and when this signaling pathway is inhibited, the protective effect of ME is abrogated. Conclusions: The use of ME is a potential therapeutic approach for regulating LIRI by activating PI3K/Akt signaling.