Linarin Protects Against CCl4-Induced Acute Liver Injury via Activating Autophagy and Inhibiting the Inflammatory Response: Involving the TLR4/MAPK/Nrf2 Pathway

Background: Linarin has been implicated in the inhibition of inflammatory responses and hepatoprotective effects. However, the precise mechanism by which Linarin integrates injury-induced signaling from inflammatory responses and oxidative stress remains unclear. Methods: We evaluated the role of Linarin in a mouse model of carbon tetrachloride (CCl 4 )-induced acute liver injury. Mice were orally pretreated with Linarin or vehicle for seven consecutive days, followed by intraperitoneal injection with 0.2% (v/v) CCl 4 . To investigate the mechanism of action on oxidative stress, CCl 4 -stimulated HepG2 cells were utilized. Results: Our results revealed Linarin remarkably attenuated the loss of hepatic architecture, inflammatory cell infiltration, serum transaminases, and pro-inflammatory cytokines induced by CCl 4 . Linarin attenuated CCl 4 -induced oxidative stress by increasing the expression of cytosolic Nrf2 (nuclear factor erythroid 2-related factor 2), inducing nuclear localization of Nrf2, and increasing stress-induced protein heme oxygenase-1 (HO-1). Additionally, Linarin decreased the expression of toll-like receptors (TLR)-4, and its downstream proteins, MyD88, IRAK1, and TRAF6. Furthermore, Linarin reversed CCl 4 -induced phosphorylation of ERK, p38, and JNK. Importantly, Linarin increased the expression of both LC3II and Beclin 1, which are hallmarks of autophagic flux. Autophagy-mediated hepatoprotective effects in Linarin-treated HepG2 cells were mitigated by the autophagy inhibitor 3-MA. However, combined treatment of Linarin with 3-MA failed to significantly reverse cell apoptosis and the production of transaminases and pro-inflammatory cytokines. Conclusion: Linarin prevents acute liver injury, possibly by alleviating ROS-induced oxidative stress, inhibiting TLR4/MyD88 and JNK/p38/ERK-mediated inflammatory responses, and promoting Beclin 1/LC3II-mediated autophagic flux. Keywords: Linarin, acute liver injury, autophagy, TLR4, MAPK, Nrf2