Stauntonia hexaphylla Extract Ameliorates Androgenic Alopecia by Inhibiting Androgen Signaling in Testosterone-induced Alopecia Mice

Figure 1.. The effects of Stauntonia hexaphylla extract on androgen signaling, apoptosis, and proliferation in human follicular dermal papilla cells (HFDPCs). Human follicular dermal papilla cells were starved for 24 h, and dihydrotestosterone (DHT) (10 nM) was treated with or without finasteride (10 nΜ) and S. hexaphylla extract (50 μM) co-treatment for 72 h. A, 5α-reductase, androgen receptor, and p-glycogen synthase kinase-3β (GSK3β); B, Relative quantitative densitometry evolution of western blot data; C, Level of dihydrotestosterone; D, The expression of Bax, Bcl-2, and poly (ADP-ribose) polymerase (PARP)-1 and relative quantitative densitometry evolution of western blot data; E, Expression of proliferating cell nuclear antigen (PCNA); F, Cell cycle; G, Cell morphology 200X. Values are expressed as mean ± SEM. *** P < 0.001, ** P < 0.01 and * P < 0.05, significant differences compared with DHT-treated cells. ### P < 0.001, significant differences compared with control cells.

Figure 2.. Analysis of morphological and histological changes caused by Stauntonia hexaphylla extract in the testosterone propionate (TP)-induced androgenic alopecia (AGA) mouse model. A, Comparison of back skin colors and hair growth in TP-induced C57BL/6 mice; B, Hair growth score; C, Comparison of histological images of hair follicles (longitudinal view, A - D (100X) transverse view, E - H (100X), I - L (400X); D, the dermis, S; subcutaneous); D, Dermal thickness; E, Quantification of cross-sectional diameters of the hair shaft/inner root sheath (IRS); F, Follicular numbers were assessed. Values are expressed as mean ± standard deviation. ** P < 0.01 and * P < 0.05, significant differences compared with the AGA group.

Figure 3.. The effects of Stauntonia hexaphylla extract on androgen signaling in the testosterone propionate (TP)-induced androgenic alopecia (AGA) mouse model. A, Dihydrotestosterone levels in the skin were determined using a commercial assay kit; B, 5α-reductase, androgen receptor, p-glycogen synthase kinase-3β (GSK3β), and β-catenin were examined; C, Relative quantitative densitometry evolution of western blot data. Values are expressed as mean ± standard deviation. ### P < 0.01, significant differences compared with control cells. *** P < 0.001 and * P < 0.05, significant differences compared with the AGA group.

Figure 4.. The effects of Stauntonia hexaphylla extract on paracrine factors, apoptosis, and proliferation in the TP-induced androgenic alopecia (AGA) mouse model. A, transforming growth factor beta-1 (TGF-β1), dickkopf-a (DKK-1), and relative quantitative densitometry evolution of western blot data; B, Immunohistochemistry of cytokeratin 14 (CK-14) in the outer root sheath, 400X (A, control; B, alopecia; C, finasteride; D, S. hexaphylla 50 mg/kg); C, Bax, Bcl-2, and the Bax/Bcl-2 ratio indicating apoptosis; D, TUNEL staining 100X (A, control; B, alopecia; C, finasteride; D, S. hexaphylla 50 mg/kg); E, Cyclin D and relative quantitative densitometry evolution of western blot data; F, Immunohistochemistry of proliferating cell nuclear antigen (PCNA) and the number of positively stained nuclei in the prostate epithelial cells. Longitudinal view (A - D, 400X); transverse view (E - H 400X) (A and E, control; B and F, alopecia; C and G, finasteride; D and H, S. hexaphylla 50 mg/kg). Values are expressed as mean ± standard deviation. ## P < 0.01 indicates significant differences compared with control cells. *** P < 0.001, ** P < 0.01 and * P < 0.05 indicate significant differences compared with…