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Deduhonghua-7 powder and its bioactive component luteolin alleviate hepatic fibrosis through CHRM3/NF-κB axis
Yumei1, Anda2, Runguo Xue3
1Clinical Medical College, Inner Mongolia Minzu University, Inner Mongolia Autonomous Region, Tongliao, 028000, China; Infectious Diseases Department, Tongliao People's Hospital, Inner Mongolia Autonomous Region, Tongliao, 028000, China.
Background:
To examine the hepatoprotective and antifibrotic properties of Deduhonghua-7 powder and its active component luteolin in a CCl4-induced liver fibrosis model, followed by in-depth analysis of the associated molecular mechanisms.
Methods:
A CCl4-induced chronic hepatic injury mouse model was constructed to evaluate the therapeutic efficacy of Deduhonghua-7 powder. Potential targets and active components were predicted through network pharmacology and molecular docking, followed by molecular dynamics simulations to verify the binding stability of luteolin to key targets. The antifibrotic mechanism of luteolin was further investigated in CCl4-stimulated LX2 human hepatic stellate cells and CHRM3-overexpression models.
Results:
Deduhonghua-7 powder significantly ameliorated CCl4-induced hepatic histopathological damage, reduced collagen deposition, and normalized serum ALT, AST, and ALP levels. Network pharmacology analysis identified 30 therapeutic targets and revealed multi-component, multi-target mechanisms. Molecular docking and dynamics studies consistently showed stable binding interactions between luteolin and core targets CHRM3 and STAT3, with binding free energies reaching ≤-13.39 kcal/mol. Notably, luteolin treatment markedly restored CHRM3 expression that was suppressed by CCl4 both in vivo and in vitro. Either CHRM3 activation via oxotremorine-M or CHRM3 overexpression significantly inhibited CCl4-induced upregulation of α-SMA and COL1A1, mimicking the antifibrotic effects of luteolin. Furthermore, luteolin-mediated CHRM3 restoration contributed to reduced hepatic inflammation, characterized by lower levels of IL-1β, IL-6, and TNF-α, and attenuated NF-κB p65 phosphorylation.
Conclusion:
Deduhonghua-7 powder exerts potent antifibrotic effects against CCl4-induced liver injury. Luteolin, a key active component, attenuates hepatic fibrosis and inflammation by upregulating CHRM3 expression, thereby inhibiting hepatic stellate cell activation and NF-κB signaling.