Ligustilide alleviates hepatic fibrosis by targeting the mesenchymal homeobox 1 (MEOX1)– transcriptional enhanced associate domain factor 2 (TEAD2) signalling axis

BACKGROUND AND PURPOSE: Mesenchyme homeobox 1 (MEOX1) has been characterised as a central transcriptional regulator of fibroblast activation. Ligustilide (LIG) has significant antifibrotic, anti-inflammatory and antioxidative activities. We investigated if LIG can ameliorate hepatic fibrosis by targeting MEOX1 and exploring the underlying mechanism. EXPERIMENTAL APPROACH: )-induced mice were used to assess the antifibrotic effects of AAV9-pGFAP-sh-MEOX1 and LIG. In vitro, LX-2 cells, following transfection with siRNA or pcDNA or pretreatment with LIG, were incubated with TGF-β1 for the assessment. KEY RESULTS: Knockdown of MEOX1 mitigated fibrosis, both in vivo and in vitro. Furthermore, through a strategy involving Traditional Chinese Medicine (TCM) prescription screening, molecular docking, cellular thermal shift assay (CETSA), site-specific mutation and surface plasmon resonance (SPR), we identified LIG as a potential inhibitor of MEOX1. Subsequent validation confirmed that LIG exerted significant antifibrotic effects through MEOX1. Mechanistic studies revealed that MEOX1 promoted transcriptional enhanced associate domain factor 2 (TEAD2) transcription, by binding to the -988 to -982 nt region of the TEAD2 promoter, which increased the transcription of Hippo signalling targets and stimulated hepatic stellate cell (HSC) activation and proliferation. LIG can also bind to the HOX domain of MEOX1, thereby inhibiting its function and alleviating hepatic fibrosis. CONCLUSION AND IMPLICATIONS: The MEOX1-TEAD2 signalling axis is crucial for hepatic fibrosis. LIG attenuates hepatic fibrosis by targeting MEOX1 and inhibiting the downstream of the Hippo signalling pathway. Our results elucidated the mechanistic basis for developing LIG as a clinical antifibrotic agent.