Work

Cannabidiol suppresses M2-like macrophage polarization via TRPV4/RhoA/ROCK signaling to alleviate hepatic fibrosis

Year: 2026

Type: article

Abstract: Background Dysregulated buildup of extracellular matrix constitutes a hallmark of hepatic fibrosis, loss of liver architectural integrity, and impaired hepatic functional capacity. Pro-fibrogenic M2-polarized macrophages promote the transdifferentiation of hepatic stellate cells and facilitate fibrotic progression. Cannabidiol (CBD) exhibits immunomodulatory and anti-fibrotic properties. However, whether CBD regulates M2-like macrophage polarization to mitigate hepatic fibrogenesis remains unclear. Objective Cannabidiol-driven mitigation of pathological hepatic remodeling was evaluated, with its molecular regulatory mechanisms systematically clarified. Methods The modulation of liver fibrosis by cannabidiol was investigated in vitro and in vivo . A model was created using bile duct ligation (BDL). Anti-fibrotic effects were tested with liver function tests, histopathology, immunohistochemistry, and immunofluorescence. Co-culture systems comprising THP-1-derived macrophages and LX-2 stellate cells were constructed to delineate cannabidiol-mediated regulation of M2-like macrophage polarization and hepatic stellate cell transactivation. Transcriptomic, single-cell transcriptomic, network pharmacology, molecular docking simulation, and dynamic conformational approaches explored cannabidiol’s binding to TRPV4. Western blot, qRT-PCR, flow cytometry, CETSA, Co-IP, and mass spectrometry detected macrophage polarization and under-healing pathway markers. Transwell and colony-formation assays assessed the effects of conditioned medium from CBD-treated M2-like macrophages on LX-2 cell migration, proliferation, and activation. Results Cannabidiol mitigated hepatic damage, fibrogenic progression, and inflammatory responses in BDL-operated mice, supported by ameliorated hepatic functional indices and reduced fibrotic biomarkers. CBD inhibited IL-4-induced M2-like polarization in THP-1 cells, resulting in decreased expression of M2-associated markers and pro-fibrotic factors. Mechanistically, CBD may engage TRPV4 and suppress RhoA/ROCK signaling, while concurrently modulating STAT6 phosphorylation. These molecular actions inhibited LX-2 cell activation, as indicated by reduced Col-I and α-SMA expression, along with diminished migration and proliferation. CBD further reduced collagen deposition and M2-associated macrophage infiltration in hepatic tissue. Notably, co-administration of CBD with a TRPV4 inhibitor potentiated anti-fibrotic effects both in vivo and in vitro .

Source: Frontiers in Immunology

DOI: 10.3389/fimmu.2026.1951332