Compound Platycladus orientalis tincture promotes hair regrowth and is associated with hair-cycle progression and Wnt/β-catenin-related signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Alopecia is traditionally managed in Chinese medicine using topical botanical preparations aimed at restoring hair-cycle dynamics and improving the follicular microenvironment. Compound Platycladus orientalis tincture (CPT), an ethanol-based polyherbal formulation consisting of Platycladi Cacumen, Szechwan Chinaberry Fruit, and Herba Speranskiae Tuberculatae, has been used clinically as an in-hospital topical preparation for hair loss, yet its chemical basis and pharmacological mechanisms remain insufficiently defined.

AIM OF THE STUDY: To characterize the chemical profile of CPT and to elucidate its efficacy, preliminary safety profile, and mechanistic basis in promoting hair regrowth in preclinical models of androgenetic alopecia (AGA) and cyclophosphamide-induced alopecia areata-like hair loss, with supporting cellular and network-level evidence.

MATERIALS AND METHODS: CPT constituents were profiled by LC-MS/MS. Putative bioactive compounds were prioritized using a multidimensional scoring framework, and candidates (TotalScore >85) were subjected to network pharmacology analysis. A dose-ranging study (0.5 × , 1 × , 2 × ) was performed in testosterone propionate-induced AGA and cyclophosphamide-induced alopecia areata-like mouse models to select the working concentration, followed by in vivo efficacy evaluation using macroscopic assessment and histopathology. In the animal experiments, mice in the Model group received topical application of the corresponding blank vehicle, whereas mice in the CPT group received CPT in the same vehicle system. Mechanistic readouts included immunohistochemistry, immunofluorescence-based spatial analysis of β-catenin and AR in skin sections, qPCR, and western blotting of key hair growth-related markers. HDPC assays were performed to evaluate proliferation, alkaline phosphatase activity, and cell-cycle progression.

RESULTS: LC-MS/MS revealed a chemically diverse profile enriched in flavonoids, polyphenols, organic acids, and terpenoids. In dose-ranging experiments, 1 × and 2 × CPT showed significant efficacy, whereas 0.5 × produced limited improvement; 1 × was therefore selected as the working concentration for downstream studies. Topical CPT significantly accelerated hair regrowth in both AGA and cyclophosphamide-induced alopecia areata-like models, increased anagen-phase follicles, reduced AR-related nuclear signal and apoptosis-related signals, increased VEGF and β-catenin, and enhanced nuclear enrichment of β-catenin in hair follicles. Transcriptional and protein analyses were consistent with involvement of Wnt/β-catenin-related signaling rather than definitive causal activation. CPT also enhanced HDPC viability, alkaline phosphatase activity, and cell-cycle progression. Network pharmacology suggested a multi-target regulatory profile associated with growth, survival, and inflammatory pathways. Under the tested dose range and observation period, CPT showed no obvious systemic toxicity based on serum ALT/AST levels and major organ histology.

CONCLUSIONS: CPT is a biologically active topical botanical formulation that promotes hair regrowth in preclinical AGA and cyclophosphamide-induced alopecia areata-like models. Its activity is associated with coordinated modulation of androgen-related and apoptosis/proliferation programs and with Wnt/β-catenin-related signaling. These findings support further mechanistic and translational investigation of CPT for alopecia management.