The Role of Ferroptosis in Liver Injury after Cold Ischemia-Reperfusion in Rats with Autologous Orthotopic Liver Transplantation

Abstract

Using autologous orthotopic liver transplantation（AOLT） model in rats, the effect of lipid reactive oxygen species（L-ROS） inhibitor Ferrostain-1 on ferroptosis signal pathway was observed to determine whether ferroptosis occurred in rat liver injury after cold Cold ischemia-reperfusion(I/R). Thirty-two healthy adult SPF male SD rats，8 ~ 10 weeks old, weight 240 ~ 260g, It is divided into four groups by the method of random number table(n = 8)：Sham group,I/R group,I/R+Fer-1 group,I/R+DFO group. In I/R+Fer-1 group, Intraperitoneal injection of ferristatin-1(5mg /kg) 30 minutes before surgery；In I/R+DFO group, DFO 100mg/kg was injected intraperitoneally 1 h before operation and 12 h after operation. Blood samples were taken from the inferior hepatic vena cava 24 hours after reperfusion, After anesthesia, the rats were killed and part of their liver tissue was removed. The pathological changes of liver tissue sections were observed under high power microscope, and the liver injury was evaluated；Determination of serum malondialdehyde (MDA) and serum levels of ALT, AST, IL-6 by ELISA method，Determination of reduced glutathione (GSH), glutathione peroxidase 4 (GPX4), MDA, Fe2+ and superoxide dismutase (SOD) in liver tissue. Compared with Sham group, the serum levels of IL-6,MDA, AST and ALT in I/R group were obviously higher (P < 0.05)；The levels of MDA and Fe2+ in liver tissue were significantly increased (P < 0.05)；The levels of SOD, GSH and GPX4 in liver tissue decreased. The levels of serum MDA, IL-6, AST and ALT in I/R+Fer-1 and I/R+DFO groups were significantly lower than those in I/R group at 24 hours after reperfusion；In I/R+Fer-1 group, the level of MDA in liver tissue decreased significantly, while the level of SOD, GSH and GPX4 in intestinal tissue increased (P < 0.05)；In I/R+DFO group, the levels of MDA and Fe2+ in liver tissue decreased significantly, while the level of SOD in intestinal tissue increased (P < 0.05).Ferroptosis is involved in pathophysiological process of liver injury after cold ischemia-reperfusion in AOLT rats.