Combined Transcriptomic and Lipidomic Profiling Reveals DHT-Associated Lipid Remodeling and Extracellular Matrix Signatures in Rabbit Hair Follicle Stem Cells

Androgen signaling is tightly linked to the initiation and progression of androgenetic alopecia (AGA). Nevertheless, the direct effects of dihydrotestosterone (DHT) on lipid metabolism and the molecular status of hair follicle stem cells (HFSCs) have not been systematically elucidated. In this study, rabbit HFSCs were stimulated with gradient concentrations of DHT in vitro. We combined cellular phenotyping, transcriptome sequencing and untargeted lipidomics to characterize DHT-induced changes, with qPCR and Western blot used for molecular validation. Under our experimental conditions, DHT induced an activated HFSC state instead of simply enhancing HFSC function; it elevated metabolic activity and clonogenic potential, increased the proportion of S-phase cells, and suppressed apoptosis without obvious acute cytotoxicity, thereby reshaping the cellular status of HFSCs. Lipidomics revealed extensive lipid-component remodeling: triglyceride (TG) species decreased, whereas sphingolipids, including sphingomyelin (SM) and hexosylceramides (Hex1Cer), were markedly enriched. Transcriptomic analysis demonstrated significant up-regulation of extracellular matrix (ECM)-related genes, including COL5A2, COL8A1, and MFAP5, accompanied by coordinated perturbation of the Wnt and TGF-β signaling pathways. Collectively, DHT induces a characteristic molecular phenotype in rabbit HFSCs characterized by altered cellular activity, lipid reprogramming, and activated ECM transcription. This work indicates that androgens do not merely suppress HFSC viability but drive complex metabolic-transcriptional remodeling, providing in vitro experimental evidence for understanding follicular pathological mechanisms in AGA.