归乌生发颗粒对雄激素性脱发小鼠毛发再生的作用与机制研究*

作者：杨晓彤1，亓智玮1，李明秀1，刘聪毅1，李雯婧1，吴丽丽1,2，刘铜华1,2，秦灵灵1,3,4，李青松5

单位：1.北京中医药大学教育部中医养生学重点实验室，北京 100029； 2.北京中医药大学中医养生研究所，北京 100029； 3.北京中医药大学科研处，北京 100029； 4.教育部高等学校学科创新引智基地，北京 100029； 5.陕西中医药大学第一临床学院，陕西 咸阳 712000

引用：引用：杨晓彤，亓智玮，李明秀，刘聪毅，李雯婧，吴丽丽，刘铜华，秦灵灵，李青松.归乌生发颗粒对雄激素性脱发小鼠毛发再生的作用与机制研究[J].中医药导报,2025,31(12):41-47.

DOI：10.13862/j.cn43-1446/r.2025.12.007

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摘要：

目的：探讨归乌生发颗粒对雄激素性脱发（AGA）模型小鼠毛发再生的效果及其机制。方法：选取60只雄性SPF级C57BL/6N小鼠，采用随机数字表法按体质量分组，分为正常组、模型组、非那雄胺组（0.13 mg/kg）、归乌生发颗粒低剂量组（0.06 g/kg）、归乌生发颗粒中剂量组（0.12g/kg）及归乌生发颗粒高剂量组（0.24 g/kg）。通过皮下多点注射二氢睾酮构建雄激素性脱发（AGA）模型，造模的同时给药，实验周期为21 d。以脱毛区域的新生毛发面积、毛发长度、毛发质量和毛囊数量为评价指标，观察归乌生发颗粒对小鼠的治疗效果；利用HE染色法观察皮肤病理变化；采用酶联免疫吸附试验（ELISA）检测皮肤中睾酮（T）、双氢睾酮（DHT）和5α-还原酶（5-AR）水平；通过免疫组化检测Wnt/β-catenin通路中Wnt10b、β-catenin蛋白及细胞凋亡相关Bcl-2、Bax蛋白的表达；使用实时荧光定量聚合酶链式反应（qRT-PCR）分析Wnt/β-catenin通路中Wnt10b、β-catenin及细胞凋亡相关Bcl-2、Bax基因的表达。结果：与正常组比较，模型组小鼠新生毛发面积、毛发长度、质量和毛囊数量均降低（P<0.05或P<0.01）。与模型组比较，各给药组上述指标均改善（P<0.05或P<0.01）。ELISA结果显示，与正常组比较，模型组小鼠T、DHT和5-AR含量升高（P<0.05或P<0.01）。与模型组比较，归乌生发颗粒低、中剂量组T、DHT水平均降低（P<0.05或P<0.01），高剂量组无显著变化（P＞0.05）。与模型组比较，各给药组5-AR含量均降低（P<0.01）。免疫组化结果表明，与正常组比较，模型组Bax蛋白表达增加，Wnt10b、β-catenin和Bcl-2蛋白表达减少（P<0.01）；与模型组比较，各给药组Bax表达显著降低，Wnt10b、β-catenin和Bcl-2表达显著增加（P<0.01）。qRT-PCR结果与免疫组化结果一致，各给药组Bax基因表达显著降低，Wnt10b、β-catenin和Bcl-2基因表达显著增加（P<0.01）。结论：归乌生发颗粒可显著改善小鼠雄激素性脱发，其可能通过降低雄激素水平、抑制5-AR、促进Wnt/β-catenin信号通路活性及抑制细胞凋亡发挥作用。

关键词：雄激素性脱发；归乌生发颗粒；Wnt/β-连环蛋白（β-catenin）信号通路；Bcl-2；Bax；小鼠

Abstract：

Objective: To evaluate the impact of Guiwu Shengfa granules on hair regeneration in mice exhibiting androgenetic alopecia (AGA) and to elucidate the associated underlying mechanisms. Methods: A total of 60 SPF male C57BL/6N mice were selected and categorized into different groups according to body weight, utilizing a random number table for allocation. The groups included normal group, model group, finasteride treatment group (0.13 mg/kg), Guiwu Shengfa granules low-dose group (0.06 g/kg), Guiwu Shengfa granules medium-dose group (0.12 g/kg), and Guiwu Shengfa granules high-dose group (0.24 g/kg). The AGA model was induced through multiple subcutaneous injections of dihydrotestosterone. The therapeutic effects of Guiwu Shengfa granules were evaluated by assessing the area, length, and weight of newly grown hair, as well as the number of hair follicles in the depilated regions. Histopathological examinations were conducted using hematoxylin and eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of testosterone (T), dihydrotestosterone (DHT), and 5α-reductase (5-AR) in skin samples. Additionally, immunohistochemistry was employed to detect the expression of Wnt10b and β-catenin proteins in the Wnt/β-catenin pathway and the apoptosis-related Bcl-2 and Bax proteins. Additionally, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was utilized to analyze the expression of Wnt10b, β-catenin, and the apoptosis-related genes Bcl-2 and Bax in the Wnt/β-catenin pathway. Results: The area, length, weight of new hair, and the number of hair follicles in the model group were significantly diminished compared to the normal group (P<0.05 or P<0.01). All treatment groups exhibited considerable improvement in these parameters compared to the model group (P<0.05 or P<0.01). ELISA data revealed that the concentrations of T, DHT and 5-AR in the model group were significantly increased compared to the normal group (P<0.05 or P<0.01). Compared to the model group, Guiwu Shengfa granules low-dose group and Guiwu Shengfa granules medium-dose group showed decreased levels of T and DHT (P<0.05 or P<0.01), while the Guiwu Shengfa granules high-dose group showed no significant changes(P＞0.05). The level of 5-AR substantially decreased in all treatment groups compared to the model group (P<0.01). Immunohistochemical results showed that compared with the normal group, the model group exhibited increased Bax protein expression and decreased expression of Wnt10b, β-catenin, and Bcl-2 proteins (P<0.01). Compared with the model group, all treatment groups showed significantly reduced Bax expression and significantly increased expression of Wnt10b, β-catenin, and Bcl-2 (P<0.01). The qRT-PCR results were consistent with the immunohistochemical findings, showing that the expression of the Bax gene was significantly decreased, while the expressions of the Wnt10b, β-catenin, and Bcl-2 genes were significantly increased in all treatment groups (P<0.01). Conclusion: Guiwu Shengfa granules can significantly ameliorate androgenetic alopecia in mice, potentially through mechanisms involving the reduction of androgen levels, inhibition of 5-AR activity, activation of the Wnt/β-catenin signaling pathway, and the suppression of apoptosis.

Key words：androgenetic alopecia; Guiwu hair growth granules; Wnt/β-catenin (β-catenin) signaling pathway; Bcl-2; Bax; mice

发布时间：2026-01-30

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