Morroniside ameliorates hepatic fibrosis in mice by targeting ACYP1 to promote NCOA4-mediated ferritinophagy and ferroptosis.

BACKGROUND

Liver fibrosis represents a pivotal histopathological stage in the progression of chronic liver disease toward cirrhosis, threatening millions of patients worldwide while therapeutic options remain scarce. Morroniside, an iridoid glycoside derived from Cornus officinalis, has been shown potent hepatoprotective properties, but its potential to attenuate liver fibrogenesis remains largely unexplored.

PURPOSE: This study aims to evaluate the therapeutic efficacy of morroniside against liver fibrosis and elucidate its underlying molecular mechanisms.

METHODS

Two distinct mouse models of liver fibrosis were established by carbon tetrachloride (CCl4)-induced and Gubra Amylin NASH (GAN) diet-induced. Mice administered morroniside at doses of 10 or 20 mg/kg by gavage concurrently with model induction. Its efficacy in improving liver fibrosis was assessed through histological analysis and immunoblotting. TGF-β-induced LX-2 cells and primary hepatic stellate cells (HSCs) were used to validate the mechanism of morroniside in promoting ferritinophagy and ferroptosis. Finally, overexpressing ACYP1 and silencing NCOA4 were applied both in vitro and in vivo to confirm the critical molecular mechanisms of morroniside in ameliorating liver fibrosis.

RESULTS

Morroniside alleviates liver fibrogenesis in both CCl4- and GAN diet-induced mouse models by targeting ACYP1, a phosphatase upregulated in fibrotic livers, thereby suppressing HSCs fibrogenic activation. Mechanistically, morroniside directly targeting ACYP1 to inhibit HERC2-mediated ubiquitination of NCOA4, thereby promoting ferritinophagy-mediated ferroptosis in HSCs. Notably, ACYP1 overexpression or NCOA4 silencing reversed the efficacy of morroniside in inducing ferritinophagy and ferroptosis in HSCs. These results establish the critical role of the ACYP1/HERC2/NCOA4 signaling axis in mediating ferritinophagy and ferroptosis, and demonstrate that targeting this pathway is a key mechanism by which morroniside suppresses liver fibrosis progression.

CONCLUSION

This study provides compelling evidence that morroniside alleviates liver fibrosis in mice by directly targeting ACYP1 to inhibit HERC2-mediated ubiquitination of NCOA4, thereby promoting ferritinophagy and ferroptosis in HSCs. These results provide new mechanistic insights into the anti-fibrotic activity of morroniside and support its further development as a potential intervention for chronic liver diseases.