[Mechanism of hyperoside in improving hepatic fibrosis via Nrf2/HO-1 and PI3K/Akt signaling pathways]

This study aims to investigate the mechanism by which hyperoside(Hyp) improves hepatic fibrosis in mice through the nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1) and phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt) signaling pathway. This study employed network pharmacology and molecular docking techniques to screen potential action targets and pathways for Hyp. Subsequently, the mechanisms of action of Hyp against hepatic fibrosis were investigated in a carbon tetrachloride(CCl_4)-induced mouse model of hepatic fibrosis and a hydrogen peroxide(H_2O_2)-stimulated human hepatic stellate cell(LX-2) model. The results of network pharmacology combined with molecular docking reveal that Hyp exhibits strong binding affinities with Nrf2, HO-1, PI3K, and Akt, suggesting that these molecules may serve as core targets for its intervention in hepatic fibrosis. In vivo experiments demonstrate that Hyp effectively reduces the levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), and γ-glutamyl transferase(γ-GT) in serum. Haematoxylin-eosin(HE), Masson's trichrome(Masson), and Van Gieson(VG) staining results reveal significant attenuation of inflammatory infiltration and collagen deposition in liver tissues. In vitro experiments demonstrate that Hyp enhances the viability of H_2O_2-injured LX-2 cells. Mechanistically, Hyp reduces levels of malondialdehyde(MDA), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-6(IL-6) levels in liver tissue and cells, while increasing superoxide dismutase(SOD) and glutathione(GSH) levels. Western blot analysis indicates that Hyp downregulates the protein expression levels of α-smooth muscle actin(α-SMA) and type Ⅰ collagen(collagen Ⅰ) as well as promotes the protein expressions of Nrf2 and HO-1, while inhibiting the phosphorylation levels of PI3K and Akt. However, when Nrf2 is knocked down in LX-2 cells and the PI3K inhibitor LY294002 is added, Hyp's antioxidant capacity is weakened while its anti-inflammatory capacity is enhanced. This study is the first to systematically reveal that Hyp exerts dual regulation of both the Nrf2/HO-1 and PI3K/Akt signaling pathways. By activating the antioxidant system and inhibiting the inflammatory response, Hyp effectively achieves the effect of anti-fibrosis. This finding elucidates the pharmacological basis of Hyp with multi-target intervention and offers a novel theoretical basis and potential targets for the prevention and treatment of hepatic fibrosis.