Supplementary file 1_Therapeutic functions of tanshinone IIA in a zebrafish model of glucocorticoid-induced osteoporosis.docx

BackgroundTanshinone IIA (TSN), a compound extracted from Salvia miltiorrhiza, has demonstrated a range of pharmacological activities. However, its therapeutic efficacy in glucocorticoid-induced osteoporosis (GIOP) remains insufficiently understood. This study was performed to investigate the protective effects of TSN against prednisolone (PN)-induced osteoporosis in zebrafish larvae.
MethodsThe osteoprotective properties of TSN were assessed using alizarin red S staining, calcein staining, and alcian blue staining to evaluate bone mineralization, density, and cartilage development. Cardiovascular function and locomotor behavior were analyzed using transgenic zebrafish models and behavioral tracking systems. RNA sequencing was performed to identify differentially expressed genes and key signaling pathways. Protein-protein interaction networks were constructed to elucidate gene associations, and results were validated via quantitative reverse transcription polymerase chain reaction.
ResultsTSN administration effectively attenuated PN-induced developmental toxicity, enhanced bone mineralization and density, and improved cartilage abnormalities in zebrafish larvae. Transcriptomic analysis identified 505 genes with reversed expression profiles between the TSN and PN treatment groups, primarily associated with skeletal system development, lipid metabolism, and fatty acid β-oxidation. Key affected pathways included PPAR signaling, lipid atherosclerosis, and reactive oxygen species. Notably, skeletal development was characterized by the upregulation of genes including col1a1b, col2a1b, col9a3, rdh1, and acana.
ConclusionThese findings demonstrate the osteoprotective effects of TSN in mitigating glucocorticoid-induced bone loss and cartilage abnormalities, highlighting its potential as a therapeutic agent for GIOP. The transcriptomic insights provide novel perspectives on TSN’s role in regulating bone metabolism and skeletal development.