Objective: To investigate the role of Maresin1 (MaR1) in hepatic ischemia-reperfusion injury (HIRI). Methods: The HIRI model was established and randomly divided into a sham operation group (Sham group), an ischemia-reperfusion group (IR group), and a MaR1 ischemia-reperfusion group (MaR1+IR group). MaR1 80ng was intravenously injected into each mouse's tail veins 0.5h before anesthesia. The left and middle hepatic lobe arteries and portal veins were opened and clamped. The blood supply was restored after 1h of ischemia. After 6h of reperfusion, the mice were sacrificed to collect blood and liver tissue samples. The Sham's group abdominal wall was only opened and closed. RAW267.4 macrophages were administered with MaR1 50ng/ml 0.5h before hypoxia, followed by hypoxia for 8h and reoxygenation for 2h, and were divided into the control group, the hypoxia-reoxygenation group (HR group), the MaR1 hypoxia-reoxygenation group (MaR1 + HR group), the Z-DEVD-FMK hypoxia-reoxygenation group (HR+Z group), the MaR1 + Z-DEVD-FMK hypoxia-reoxygenation group (MaR1 + HR + Z group), and the Con group without any treatment. Cells and the supernatant above them were collected. One-way analysis of variance was used for inter-group comparisons, and the LSD-t test was used for pairwise comparisons. Results: Compared with the Sham group, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin (IL)-1β, and IL-18 in the IR group were significantly higher (P < 0.05), with remarkable pathological changes, while the level in the MaR1 + IR group was lower than before (P < 0.05), and the pathological changes were alleviated. Compared with the Con group, the HR group had higher levels of IL-1β and IL-18 (P < 0.05), while the MaR1 + HR group had lower levels of IL-1β and IL-18 (P < 0.05). Western blot showed that the expressions of caspase-3, GSDME, and GSDME-N were significantly higher in the HR group and IR group than in the other groups; however, the expression was lower following MaR1 pretreatment. The Z-DEVD-FMK exploration mechanism was inhibited by the expression of caspase-3 in HIRI when using MaR1. Compared with the HR group, the IL-1β and IL-18 levels and the expressions of caspase-3, GSDME, and GSDME-N in the HR + Z group were decreased (P < 0.05), while the expression of nuclear factor κB was increased, but following MaR1 pretreatment, nuclear factor κB was decreased. There was no significant difference in the results between the MaR1 + H/R group and the MaR1 + H/R + Z group (P > 0.05). Conclusion: MaR1 alleviates HIRI by inhibiting NF-κB activation and caspase-3/GSDME-mediated inflammatory responses.
目的： 探讨Maresin 1（MaR1）在肝缺血再灌注损伤（HIRI）中的作用。 方法： 建立HIRI模型，随机分为假手术组（Sham组）、缺血再灌注组（IR组）、MaR1缺血再灌注组（MaR1+IR组）。在麻醉前0.5 h尾静脉注射MaR1 80 ng/只，打开腹腔夹闭左、中肝叶动脉和门静脉，缺血1h后恢复供血，再灌注6 h后处死小鼠收集血液、肝组织，Sham组仅打开、关闭腹腔。RAW267.4巨噬细胞在缺氧前0.5 h给予MaR1 50 ng/ml，行缺氧8 h复氧2 h，分为对照组（Con组）、缺氧复氧组（HR组）、MaR1缺氧复氧组（MaR1+HR组）、Z-DEVD-FMK缺氧复氧组（HR+Z组）、MaR1+Z-DEVD-FMK缺氧复氧组（MaR1+HR+Z组），Con组未做任何处理，收集细胞和上清液。组间比较采用单因素方差分析，其中两两比较采用LSD-t检验。 结果： 与Sham组相比，IR组丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、白细胞介素（IL）-1β、IL-18水平明显升高（P<0.05），病理改变明显；MaR1+IR组水平较前降低（P<0.05）、病理改变减轻。与Con组比较，HR组IL-1β和IL-18水平升高（P<0.05），MaR1+HR组IL-1β和IL-18水平较前降低（P<0.05）。蛋白质印迹法检测胱天蛋白酶（caspase）-3、焦孔素（GSDM）E、GSDME-N蛋白的表达，HR组和IR组明显高于其他组，经MaR1预处理后表达降低。为了探究MaR1在HIRI中的机制，用Z-DEVD-FMK抑制caspase-3的表达。与HR组比较，HR+Z组IL-1β和IL-18水平降低（P<0.05），caspase-3、GSDME、GSDME-N表达减少、核因子κB表达增加，经MaR1预处理后核因子κB表达降低。MaR1+HR组与MaR1+HR+Z组比较，结果差异无统计学意义（P > 0.05）。 结论： MaR1通过抑制核因子κB活化、caspase-3/GSDME介导的炎性反应，减轻HIRI。.
Keywords: Caspase-3; Ischemia-reperfusion injury, liver; Maresin 1; Pyroptosis.