Study on the role of cyanidin-3‑O‑galactoside in alleviating pulmonary fibrosis via Ccl3/Slc2a3

Cyanidin-3-O-galactoside (C3G) has a variety of biological activities. Pulmonary fibrosis (PF) is a fatal interstitial lung disease. To define its pathogenic networks, we performed transcriptomic sequencing of lung tissues from five mouse groups: WT_Control, WT_Model, WT_Model_C3G, Control_Ccl3-/-, Model_Ccl3-/-. Comprehensive assessment of immune infiltration, weighted gene co-expression network, and functional enrichment verified that PF is tightly linked to immune microenvironment dysregulation. Seven machine learning algorithms identified five core immune-related targets: Ccl3, Xcl1, Pyy, Il31ra, Ppbp. Subsequent analysis prioritized Ccl3 and Ppbp as the key core pathogenic genes. Molecular docking clarified their binding modes and key interaction sites with C3G. Histopathological and biochemical assessments showed that the WT_Model group exhibited elevated expressions of Ccl3 and Ppbp, accompanied by marked inflammatory infiltration and collagen deposition. C3G intervention significantly ameliorated these PF phenotypes. Ccl3 knockout improved survival and ameliorated fibrotic pathology. Interaction modeling and correlation analyses identified Slc2a3 as a key Ccl3 downstream target. Western blot (WB) and immunohistochemical validation revealed reduced expression of Ccl3, Slc2a3, and α-SMA proteins in Model_Ccl3-/- compared to WT_Model, which suggested Ccl3 may positively regulate the expression of Slc2a3. Through transcriptome and external metabolome verification, it was found that Slc2a3 can regulate PF through metabolic pathways. In conclusion, this study confirmed the potential role of C3G in regulating Ccl3-related inflammation and metabolic pathways in bleomycin-induced PF, which is expected to become a new strategy for targeted therapy of PF and provide a theoretical basis for the development of clinical treatment.