Study on therapeutic effect of anemoside B4 on mice with androgenetic alopecia and its mechanism of action

LIU Xiaoyan; WANG Qinqin; GAO Xiang; XIE Xiaoyun; CHEN Xiate; YUAN Renyikun; GAO Hongwei; YANG Shilin

doi:10.3969/j.issn.1674-3806.2025.11.12

Abstract：Objective To explore the therapeutic effect of anemoside B4(AB4)on androgenetic alopecia(AGA)in mice and its mechanism of action.Methods Forty-eight male C57BL/6 mice were selected and randomly divided into control group,model group,low-dose AB4 group,medium-dose AB4 group,high-dose AB4 group and 5％minoxidil tincture group,with 8 mice in each group.The mice in the control group were fed normally without special interventions.The model group and each intervention group were topically applied with testosterone solution[10 mg/(kg·d)]on the skin on the neck and back of the mice to establish the AGA model.After construction of the AGA model,the model group was topically applied with 20％glycerol solution on the skin on the neck and back of the mice,twice daily,0.2 mL per time,and the low-dose AB4 group,the medium-dose AB4 group and the high-dose AB4 group were topically applied with 5 mg/mL,10 mg/mL and 20 mg/mL AB4 solution on the skin on the neck and back of the mice,respectively,twice daily,0.2 mL per time,and the 5％minoxidil tincture group was topically applied with 5％minoxidil tincture on the skin on the neck and back of the mice,twice daily,0.1 mL per time.The interventions lasted for 23 days,during which testosterone solution[5 mg/(kg·d)]was topically applied on the skin on the neck and back of the mice to maintain the AGA model.The hair growth of the mice was observed and recorded by taking photos every week,and skin hair follicles were observed by using hematoxylin and eosin(HE)staining after the last intervention.The serum testosterone,estradiol(E2)and type Ⅱ 5-α reductase(5 AR2)levels were detected by using enzyme-linked immunosorbent assay(ELISA)and the testosterone/E2 ratio was calculated.The levels of transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1 β)and interleukin-6(IL-6)in the skin tissues of the neck and back of the mice were detected.Results After 23 days of interventions,compared with the model group,the number of skin hair follicles in the AB4 intervention groups was significantly increased(P＜0.05),with larger hair follicle volume.Moreover,the number of hair follicles in the medium-dose AB4 group and the high-dose AB4 group was larger than that in the 5％minoxidil tincture group(P＜0.05).The levels of serum testosterone,5AR2 and testosterone/E2 ratio in the AB4 intervention groups were significantly lower than those in the model group(P＜0.05),while the levels of E2 in the AB4 intervention groups were significantly higher than those in the model group(P＜0.05).The level of serum E2 in the high-dose AB4 group was higher than that in the 5％minoxidil tincture group,and the testosterone/E2 ratio in the high-dose AB4 group was lower than that in the 5％minoxidil tincture group,with statistically significant differences(P＜0.05).The levels of TGF-β1 and IL-6 in skin tissues of the AB4 intervention groups were significantly lower than those in the model group and the 5％minoxidil tincture group(P＜0.05).Conclusion AB4 may exert therapeutic effect on AGA by regulating serum hormone balance and inhibiting skin inflammatory response.