Partial Liver Ischemia-Reperfusion Injury Model

Partial (70%) liver warm ischemia mouse model was established as previously described (Chen et al., 2021 (link)). Briefly, mice were first anesthetized by pentobarbital sodium (60 mg/kg; Sigma) and subjected to midline laparotomy. A microvascular clip was used to clamp the left and middle portal vein and hepatic artery branches to interrupt the blood supply of the liver. After 1 h of ischemia, the clamp was removed for reperfusion. After 6 h reperfusion, the animals were sacrificed to collect liver and serum samples for further analysis. The residual blood was discharged by portal vein injection of normal saline. Part of the liver tissue was stored in liquid nitrogen and then transferred to the refrigerator at −80°C. Another part of liver tissue was preserved in 10% formalin for pathological examination. As a sham control group, mice underwent the same surgical procedure but without vasculature clamping. To inhibit MIF and ASK1 in mice, specific ASK1 inhibitor NQDI-1 (Sigma; 10 mg/kg, dissolution by DMSO and dilute with PBS to 1.25 mg/ml, 200 μl per mice) and MIF inhibitor ISO-1 (MedChemExpress LLC; 3.5 mg/kg, dissolution by DMSO and dilute with PBS to 0.44 mg/ml, 200 μl per mice) were intraperitoneally injected 2 h before the ischemic surgery. The same volume of DMSO (diluted to 0.1% by PBS) was used as control (Xu et al., 2020 (link); Liu et al., 2021 (link)).