Dahuang-Gancao Decoction Ameliorates Testosterone-Induced Androgenetic Alopecia in Mice

Huanzhu Qian; Zhen Ye; Yu Hu; Liulin Chen; Linzhen Li; Kaihua Qin; Qiaobo Ye; Xiaohong Zuo

doi:10.1016/j.jep.2025.119347

Dahuang-Gancao Decoction Ameliorates Testosterone-Induced Androgenetic Alopecia in Mice

J Ethnopharmacol. 2025 Jan 10:119347. doi: 10.1016/j.jep.2025.119347. Online ahead of print.

Authors

Huanzhu Qian¹, Zhen Ye¹, Yu Hu¹, Liulin Chen¹, Linzhen Li¹, Kaihua Qin², Qiaobo Ye³, Xiaohong Zuo⁴

Affiliations

¹ School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
² Health Preservation and Rehabilitation College, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
³ School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, China. Electronic address: yeqiaobo@cdutcm.edu.cn.
⁴ Eye School of Chengdu University of Traditional Chinese Medicine. Ineye Hospital of Chengdu University of Traditional Chinese Medicine, KeyLaboratory of Sichuan Province Ophthalmopathy Prevention & Cureand Visual Function Protection with Traditional Chinese Medicine Laboratory. Electronic address: zuoxh0806@cdutcm.edu.cn.

PMID: 39800247
DOI: 10.1016/j.jep.2025.119347

Abstract

Ethnopharmacological relevance: Dahuang-Gancao decoction (DGD) is a traditional Chinese medicinal formula that is recorded in the Synopsis of the Golden Chamber, and is widely used to treat damp-heat in the body. Since the pathological factors of androgenetic alopecia (AGA) also reflect damp-heat blockage, DGD has great potential for the treatment of AGA and has been used effectively in clinical practice.

Aim of the study: The aim of the study was to investigate whether external application of DGD could promote the activation and proliferation of hair follicle stem cells (HFSCs) and improve AGA through the Wnt/β-catenin pathway.

Materials and methods: The main chemical components of DGD-contained serum were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database search. Cell Counting Kit-8 (CCK8) was used to investigate the appropriate concentration. Hair regeneration was assessed by hair growth score and histopathological staining. The proliferation of HFSCs and the activation of Wnt/β-catenin pathway were detected by western blot, immunofluorescence staining, real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA). The AGA mouse model was induced by external application of testosterone (T). Immunofluorescence staining was performed to localize HFSCs by CK15, followed by staining with Ki67, β-catenin, and Cyclin D1, respectively.

Results: The results illustrated that the 10% DGD group and the 10% DGD + HLY78 group could significantly promote the expression of Wnt10b and β-catenin and the proliferation of HFSCs in vitro, while the 10% DGD + IWR-1 group could reverse the promotion effect of DGD. Animal experiments showed that compared with the model group (T group), DGD promoted hair follicles to enter the anagen phase, as evidenced by an increase in hair growth score, an increase in the number of hair follicles in hematoxylin and eosin (HE) staining, and a significant increase in the ratio of the number of anagen follicles to the total number of hair follicles (AF/AF + TF). In addition, DGD upregulated the expression of Wnt/β-catenin signaling pathway proteins in the skin tissues of AGA mice. It also promoted the proliferation of HFSCs and the expression of β-catenin and Cyclin D1 cytokines in the region of HFSCs.

Conclusion: Both oral and external application of DGD can promote the proliferation of HFSCs by activating the Wnt/β-catenin signalling pathway. External application of DGD can promote the hair follicles to enter the anagen phase, which can ameliorate the symptoms of alopecia in AGA mice. Therefore, compared to oral DGD, external application of DGD is an effective and safer way of administration for the treatment of AGA.

Keywords: Androgenetic alopecia; Dahuang-Gancao decoction; Hair follicle stem cell; Wnt/β-catenin.