目的 基于经皮吸收成分探究中药育发凝胶对雄激素性脱发（androgenetic alopecia，AGA）小鼠的药效物质基础及作用机制。方法 采用涂抹丙酸睾酮建立AGA小鼠模型，涂抹中药育发凝胶进行治疗，观察小鼠毛发的生长情况；苏木素-伊红（hematoxylin-eosin，HE）染色观察毛囊状态和真皮厚度； ELISA检测小鼠皮肤组织中转化生长因子-β1（transforming growthfactor-β1，TGF-β1）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和白细胞介素-6（interleukin-6，IL-6）的含量。UPLCQTOF-MS定性分析中药育发凝胶提取液化学成分、经皮入血成分和皮肤滞留成分。采用网络药理学和分子对接预测中药育发凝胶治疗AGA小鼠的潜在作用机制；采用Western blotting检测小鼠皮肤组织中雄激素受体（androgen receptor，AR）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）和Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）的表达，验证网络药理学预测的关键靶点。结果 与对照组比较，模型组小鼠毛囊数减少且毛发生长缓慢，说明模型建立成功；与模型组比较，中药育发凝胶高剂量组小鼠新生毛发区毛囊和真皮厚度增加（P＜0.01、0.001），皮肤组织中TGF-β1、TNF-α和IL-6水平显著降低（P＜0.05、0.001）。共鉴定出中药育发凝胶提取液中76种成分，皮肤滞留成分30种，经皮入血成分7种；基于经皮吸收成分共得到成分靶点481个、疾病靶点236个、共同靶点41个，核心靶点包括蛋白激酶B（protein kinase B1，AKT1）、连环蛋白1（catenin beta 1，CTNNB1）、TNF等，基因本体（gene ontology，GO）功能富集分析共375个条目，京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集通路共117条。与模型组比较，中药育发凝胶组小鼠皮肤组织中AR、Caspase-3、Bax蛋白表达水平显著降低（P＜0.05、0.001），Bax/Bcl-2值显著降低（P＜0.001）。结论 初步揭示了中药育发凝胶对AGA小鼠的经皮吸收成分和作用机制，为其临床应用提供了理论参考。

Objective To explore the pharmacodynamic material basis and mechanism of Yufa Gel (中药育发凝胶) on androgenic alopecia (AGA) mice based on percutaneous absorption of ingredients. Methods The AGA mouse model was established by smearing testosterone propionate, and the growth of mouse hair was observed after smearing Yufa Gel; Hematoxylin-eosin (HE) staining was used to observe the status of hair follicles and the thickness of dermis; ELISA was used to detect the contents of transforming growth factor-β1 (TGF-β1), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in skin tissue of mice. UPLC-QTOF-MS was used to qualitatively analyze the chemical components of Yufa Gel extract, transdermal components and skin retention components. Network pharmacology and molecular docking were used to predict the potential mechanism of Yufa Gel in treating AGA mice; Western blotting was used detect the expressions of androgen receptor (AR), cysteine aspartate protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in skin tissue of mice to validate key targets predicted by network pharmacology. Results Compared with control group, mice in model group showed a decrease in the number of hair follicles and slow hair growth, indicating the successful establishment of model; Compared with model group, the thickness of hair follicle and dermis in new hair area of mice in Yufa Gel high-dose group was increased (P < 0.01, 0.001), and the levels of TGF-β1, TNF-α and IL-6 in skin tissue were significantly reduced (P < 0.05, 0.001). A total of 76 chemical components were identified in Yufa Gel extract, 30 skin retention components and seven transdermal blood components were analyzed; Based on percutaneous absorption of components, a total of 481 component targets, 236 disease targets, and 41 common targets were obtained. The core targets included protein kinase B1 (AKT1), catenin beta 1 (CTNNB1), TNF, etc. Gene ontology (GO) function enrichment analysis included 375 entries, and Kyoto encyclopedia of genes and genomes (KEGG) enriched pathway included 117 entries. Compared with model group, the expression levels of AR, Caspase-3 and Bax protein in skin tissue of mice in Yufa Gel group were significantly down-regulated (P < 0.05, 0.001), and Bax/Bcl-2 value was significantly decreased (P < 0.001). Conclusion The composition and mechanism of Yufa Gel on percutaneous absorption of AGA mice are preliminarily revealed, which provides a theoretical reference for its clinical application.

R285.5

天津市技术创新引导专项——企业科技特派员项目（23YDTPJC00590）