Artemisinin-hydroxychloroquine combination from traditional antimalarial medicine attenuates pulmonary fibrosis via PI3K/AKT/GSK3β-mediated inhibition of epithelial-mesenchymal transition.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisinin and hydroxychloroquine, derived respectively from Artemisia annua L. (Qinghao) and Cinchona calisaya Wedd., have been widely used for centuries to treat malaria and inflammatory disorders. In addition to their antimalarial activity, these compounds exhibit multifunctionality, including anti-inflammatory, antifibrotic, and immunomodulatory effects. Although preliminary studies suggested that the artemisinin-hydroxychloroquine (AH) combination may attenuate idiopathic pulmonary fibrosis (IPF), its underlying mechanisms remain poorly understood.

AIM OF THE STUDY: This study aimed to systematically investigate the antifibrotic efficacy of AH and its underlying molecular mechanisms through in vivo and in vitro experiments.

MATERIALS AND

METHODS

IPF was induced in Sprague-Dawley rats via intratracheal bleomycin (BLM) instillation. Beginning on day 7 post-modeling, animals received oral AH or nintedanib. Lung structure and fibrosis severity were assessed by small animal energy spectrum CT imaging, pulmonary function tests, hematoxylin-eosin (HE) and Masson's trichrome staining, and lung tissue hydroxyproline (HYP) quantification. The mechanism of AH was examined by Western blot, immunohistochemistry, and transcriptomic analysis. An in vitro epithelial-mesenchymal transition (EMT) model was established in A549 cells via TGF-β1 induction to validate AH's effects.

RESULTS

AH treatment produced multidimensional improvements in BLM-induced pulmonary fibrosis, effectively alleviating pathological tissue damage and respiratory dysfunction, reducing and mitigating pulmonary inflammation and collagen deposition. Mechanistically, our integrated transcriptomic analysis and experimental validation revealed that AH undefined anti-pulmonary fibrosis effects modulate the PI3K/AKT/GSK3β signaling pathway to inhibit epithelial-mesenchymal transition, thereby slowing the progression of idiopathic pulmonary fibrosis.

CONCLUSION

Our study provides the first evidence that the traditional antimalarial combination AH alleviates IPF by specifically modulating the PI3K/AKT/GSK3β signaling axis to inhibit EMT. These findings reveal a novel drug repurposing opportunity and underscore the potential of AH as a promising therapeutic candidate for IPF.