Rosmarinic acid alleviates acetaminophen-induced hepatotoxicity by targeting Nrf2 and NEK7-NLRP3 signaling pathway.

Rosmarinic acid (RA) is a natural polyphenol with various biological activities, such as anti-oxidative, anti-fibrotic, and hepatoprotective properties. The objective of this study was to investigate the protective effect of RA against acetaminophen (APAP)-induced hepatotoxicity (AILI) and explore the underlying mechanisms. Kunming mice were treated with RA (20, 40, or 80 mg/kg, i.g) for 7d, followed by an intraperitoneal injection of APAP (500 mg/kg). The liver injury was evaluated by serum biochemical and liver histopathological examinations. Human HepG cells were pre-treated with RA (20, 40, or 80 μmol/L) and then incubated with APAP (25 mmol/L) for 24 h. The MTT assay, wound healing assay, transwell migration assay, flow cytometry, and western blotting were employed to further evaluate RA's protective effects on AILI and explore the mechanisms. The results indicated that RA pre-treatment lowered the serum ALT and AST levels, ameliorated the histological damage to the liver, and reduced ROS generation and the production of IL-1β and IL-18 in the liver tissues in APAP-treated mice. Moreover, pre-treatment with RA could promote the cell viability and migration ability and inhibit apoptosis in APAP-treated HepG2 cells. Mechanistically, RA could significantly suppress the APAP-induced activation of the NEK7-NLRP3 signaling pathway. Notably, depletion of Nrf2 by short hairpin RNA (shRNA) partly eliminated the protective effects of RA on AILI and the suppression of NEK7-NLRP3 signaling by RA. In summary, these results indicate that RA has a protective role against AILI through Nrf2-mediated inhibition of ROS production and suppression of the NEK7-NLRP3 pathway.