Licochalcone B attenuates pulmonary fibrosis via inhibiting ZBP1-dependent PANoptosis

Pulmonary fibrosis (PF) is a chronic, progressive, and frequently fatal interstitial lung disease. Glycyrrhiza uralensis Fisch, a traditional Chinese medicine (TCM) herb, has long been used for respiratory disorders due to its anti-inflammatory and expectorant properties. Licochalcone B (LCB), a chalcone derivative isolated from Glycyrrhiza uralensis Fisch, shows therapeutic potential in airway inflammation and alveolar injury, making it a promising candidate for fibrotic lung diseases.

This study evaluates the anti-fibrotic efficacy of LCB against PF progression and elucidates its potential molecular mechanisms.

PF was induced in mice by intratracheal administration of bleomycin (BLM, 1.25 mg/kg). After disease model induction, the mice were treated with LCB (3.13 or 6.25 mg/kg per day) or pirfenidone (PFD, 300 mg/kg per day) for 14 days. Histopathological changes, collagen deposition, fibrosis-related factor levels, and PANoptotic marker protein in lung tissue were evaluated. Two TGF-β1-induced PF cell models (NIH-3T3 and BEAS-2B) were utilized to simulate fibroblast activation and epithelial-mesenchymal transition processes. The effects of LCB intervention on cell proliferation, migration, and the expression of vimentin, fibronectin (FN), and type I collagen (COL1A1) were assessed through wound healing assays, colony formation assays, Western blotting (WB), and immunocytochemistry.

LCB significantly alleviates BLM-induced pulmonary inflammation and fibrosis, reduces collagen deposition in lung tissues of fibrotic mice, and downregulates FN expression while upregulating E-cadherin (E-Cad) levels. Immunohistochemical analysis revealed that LCB downregulates the expression of ZBP1 and apoptotic marker proteins in the lung tissues of PF mice. Additionally, LCB inhibits TGF-β1-induced abnormal migration of BEAS-2B cells and aberrant proliferation of NIH/3T3 cells while suppressing the expression of fibrosis-related factors, including COL1A1, FN, and α-smooth muscle actin (α-SMA). Our findings demonstrate that ZBP1 overexpression in epithelial cells attenuates the anti-fibrotic efficacy of LCB through the activation of PANoptosis and identify the inhibition of IRF1 binding to the ZBP1 promoter as a pivotal mechanism underlying the therapeutic potential of LCB in PF.

This study found that LCB exerts pleiotropic antifibrotic effects by targeting ZBP1-mediated PANoptosis, thereby identifying ZBP1 as a critical therapeutic target for PF and highlighting LCB's potential in anti-fibrotic therapy.