(+)-Cholesten-3-One Promotes Proliferation of Hair Follicle Stem Cells via Wnt/β-Catenin Pathway

Hair follicle stem cells (HFSCs) possess the ability to repair hair follicles and the epidermis. Plastrum testudinis is used to treat skin wounds. However, it is unclear whether (+)-cholesten-3-one, an important steroid extracted from plastrum testudinis, could promote HFSCs to repair skin wound. In this study, HFSCs were isolated from hair follicles of rat vibrissae and identified by immunofluorescence. Proliferation of HFSCs treated with (+)-cholesten-3-one was tested using the CCK-8 assay. Wnt/beta-catenin expression in HFSCs stimulated with (+)-cholesten-3-one was analyzed by immunofluorescence, western blotting, and RT-qPCR. In vivo, HFSCs were injected into the dermis of rat vibrissae near a skin defect to detect whether (+)-cholesten-3-one promoted HFSCs to repair the skin wound. The results show that HFSCs were typically cobblestone-like and positive for CD34 and cytokeratin 15. (+)-Cholesten-3-one induced proliferation of HFSCs; upregulation of beta-catenin, Lef1, c-Myc, and cyclin D1 in HFSCs; and downregulation of GSK-3 beta and Tcf3. On the 4th day after transplantation of HFSCs, (+)-cholesten-3-one inhibited inflammatory reactions, and the newly formed epidermis was similar to that of normal skin. In conclusion, (+)-cholesten-3-one activated the Wnt/beta-catenin pathway and accelerated HFSCs to repair skin wound. This compound may offer a new approach for stem cell-based treatment besides conventional therapy.