Levistilide A alleviates pulmonary fibrosis by blocking the cation channel PIEZO1-centered signaling pathway in fibroblasts.

Background

Pulmonary fibrosis (PF) is a common pathological feature of various chronic lung diseases, characterized by destruction of lung parenchyma and alveolar structure, aberrant angiogenesis and fibroblast proliferation. Therefore, determining the main contributing factors of PF and finding effective targeted medications are an urgent concern that needs to be resolved. This study investigated the pharmacological mechanisms on therapeutic effects of levistilide A (LevA) for PF.

Methods

Bleomycin (BLM)-induced PF mice and mice fibroblast cell line L929 cells were used to investigate the inhibitory effect of LevA on calcium signaling-mediated angiogenesis and fibroblast growth, and explain its underlying mechanism by the RNA-sequencing analysis, the protein-protein interactions and the gene functional correlation analysis, as well as molecular biology experiments.

Results

We first demonstrated that PIEZO1 was enhanced in the lungs of PF patients and mouse models and strongly associated with angiogenesis and fibroblast proliferation. Notably, LevA remarkably improved BLM-induced PF by suppressing the cation channel PIEZO1 and calcium signaling in fibroblasts, which reduced the CAMKK2/CAMK1/p-CREB/HIF-1α/VEGF pathway-mediated angiogenesis, as well as the CALPAIN2/WNT/β-CATENIN pathway-linked fibroblast growth. According to our in vitro and in vivo studies, the continuous activation of PIEZO1 counteracted the therapeutic benefits of LevA. Mechanically, LevA directly bound to and promoted K63-linked polyubiquitination and subsequent degradation of PIEZO1 in fibroblasts via interaction with the E3 ligase TRIM25.

Conclusion

LevA ameliorates PF via inhibiting PIEZO1 and calcium signaling-centered angiogenesis and fibroblast proliferation pathways. This study provides a new scientific foundation and perspective for PF therapy.