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Abelmoschus manihot Polysaccharides Target the AMPK/Nrf2/HO-1 Signalling Pathway to Inhibit Macrophage M1
Shixing Li1, Zukuan Chang1, Jinzhan Liu1
1Department of Interventional Therapy, Xinxiang Central Hospital, Xinxiang, Henan Province, China.
Abstract:
Liver fibrosis (LF) is a critical pathological stage in the progression from chronic liver disease to cirrhosis. While Abelmoschus manihot polysaccharides (AMP) demonstrate hepatoprotective potential, the underlying anti-fibrotic mechanisms remain incompletely understood. A carbon tetrachloride-induced LF mouse model was established followed by AMP intervention. Comprehensive assessments included ELISA measurement of serum biochemical indicators (ALT, AST, ALP, TBA, TBil), fibrosis markers (HA, PC III, LN, Hyp) and inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-10). Liver tissue pathological changes were evaluated through haematoxylin-eosin (HE), Masson and Sirius Red staining. In vitro experiments utilised TGF-β1-stimulated JS-1 and LPS/IFN-γ-induced M1-type RAW264.7 macrophages. Experimental techniques encompassed MTT assay, immunofluorescence, qPCR, Western blot and flow cytometry to analyse cellular markers and signalling pathways. The study specifically investigated AMP's regulatory effects on macrophage polarisation and hepatic stellate cell activation, using the AMPK inhibitor (Compound C) to explore potential mechanistic pathways. AMP treatment effectively mitigated CCl4-induced liver injury by reducing serum markers (ALT, AST, HA, PC III, Hyp) and pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) while increasing anti-inflammatory IL-10. Mechanistically, AMP regulated macrophage polarisation (inhibiting M1 markers iNOS/CD80/CD86 while promoting M2 markers CD206/Arg-1) and suppressed fibrogenesis by downregulating α-SMA, Collagen I and TIMP1 while upregulating MMP13. These effects were mediated through activation of the AMPK/Nrf2/HO-1 pathway, ultimately inhibiting HSCs activation and extracellular matrix (ECM) accumulation. The AMPK inhibitor Compound C attenuates these protective effects. AMP alleviates LF by activating the AMPK/Nrf2/HO-1 pathway, modulating macrophage polarisation and ECM remodelling, demonstrating promising therapeutic potential.
Insights
Abelmoschus manihot polysaccharides (AMP) show potential in treating liver fibrosis (LF) by activating the AMPK/Nrf2/HO-1 pathway. This natural compound modulates macrophage polarization and reduces extracellular matrix accumulation, offering therapeutic promise.
Area of Science:
- Pharmacology
- Hepatology
- Immunology
Background:
- Liver fibrosis (LF) is a severe condition preceding cirrhosis.
- Abelmoschus manihot polysaccharides (AMP) have shown hepatoprotective effects, but their anti-fibrotic mechanisms require elucidation.
- Understanding these mechanisms is crucial for developing new LF therapies.
Purpose of the Study:
- To investigate the anti-fibrotic mechanisms of AMP in a carbon tetrachloride-induced LF mouse model.
- To explore AMP's effects on macrophage polarization and hepatic stellate cell (HSC) activation.
- To identify the key signaling pathways involved in AMP's therapeutic action.
Main Methods:
- Established a carbon tetrachloride-induced LF mouse model and treated with AMP.
- Assessed serum biochemical and fibrosis markers using ELISA.
- Performed histological analyses (HE, Masson, Sirius Red) on liver tissues.
- Conducted in vitro studies on JS-1 cells and RAW264.7 macrophages using MTT, immunofluorescence, qPCR, Western blot, and flow cytometry.
- Investigated the role of the AMPK/Nrf2/HO-1 pathway and used an AMPK inhibitor (Compound C).
Main Results:
- AMP treatment significantly reduced liver injury markers (ALT, AST) and fibrosis indicators (HA, PC III, Hyp).
- AMP suppressed pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) and increased anti-inflammatory IL-10.
- AMP modulated macrophage polarization from M1 to M2 phenotype and inhibited HSC activation by downregulating α-SMA, Collagen I, and TIMP1, while upregulating MMP13.
- These effects were linked to the activation of the AMPK/Nrf2/HO-1 pathway, as evidenced by Compound C's attenuation of AMP's benefits.
Conclusions:
- AMP effectively alleviates carbon tetrachloride-induced liver fibrosis.
- AMP exerts its therapeutic effects by activating the AMPK/Nrf2/HO-1 pathway.
- AMP modulates macrophage polarization and inhibits HSC activation and extracellular matrix accumulation, highlighting its potential as a novel anti-fibrotic agent.
