Senkyunolide I attenuates hepatic ischemia/reperfusion injury in mice via anti-oxidative, anti-inflammatory and anti-apoptotic pathways

Abstract

Background: Senkyunolide I (SEI)exerts considerable protective effects in various disease models, but its effect on hepatic ischemia-reperfusion (I/R) injury remains unknown. This research aimed to investigate the effect of SEI in a murine model of hepatic I/R injury.

Methods: With modified liver I/R murine model, low, medium and high doses of SEI were injected intraperitoneally after operation. After 6 h of reperfusion, the blood and liver were collected. Serum ALT and AST were detected by automatic analyzer, while liver injury was evaluated by HE staining. High-dose SEI was selected to further explore its impacts on oxidative stress, inflammatory responses and apoptosis induced by hepatic I/R. The pharmacological effect of SEI was also compared with a positive control, glutathione (GSH). We used ELISA to detect serum TNF-α, IL-1 β and IL-6, special kit to explore activities of SOD and GSH-Px, and the content of MDA, and western blotting to detect HO-1, Bax and Bcl-2 levels, and to perceive expressions and phosphorylations of NF- κB p65 and p38/ERK/JNK in liver tissues. Apoptosis in liver tissue was evaluated by TUNEL. The antioxidative effect of SEI was further investigated using the HuCCT1 cells stimulated with H₂O₂ and the role of SEI on regulation of Nrf-2/HO-1 was determined.

Results: 200 mg/kg of SEI was optimal dose for treating liver I/R injury. Elevated ALT, AST and histopathological injury in I/R liver was attenuated by SEI administration, similarly to GSH. Serum TNF-α, IL-1β, and IL-6 were reduced in liver I/R mice treated with SEI, and in liver tissues, phosphorylation of p65 NF-κB and MAPK kinases (p38, ERK, JNK), were inhibited. SEI reduced the MDA content, but increased HO-1 level and enhanced SOD and GSH-Px activities. Apoptosis of liver tissues was decreased, while SEI inhibited Bax and elevated Bcl-2 expression. In in vitro experiments, H₂O₂ reduced the survival rate of HuCCT1 cells, which was protected by SEI administration. SEI reduced the ROS and MDA content. The transportation of Nrf-2 into the nucleus was enhanced and HO-1 expression was upregulated.

Conclusions: SEI attenuates hepatic I/R injury in mice via anti-oxidative, anti-inflammatory and anti-apoptotic pathways.

Keywords: Apoptosis; Inflammatory response; Ischemia-reperfusion; Liver injury; Oxidative stress; Senkyunolide I.