Kim Hwapyung, Kim Gwangpyung, Kim Namsoo Peter, Kim Boyong
Department of Plant Medicals, Gyeongkuk National University, Andong 36729, Republic of Korea.
Department of Biological Science, Gyeongkuk National University, Andong 36729, Republic of Korea.
Pharmaceuticals (Basel). 2025 Aug 13;18(8):1192. doi: 10.3390/ph18081192.
Androgenetic alopecia suppresses hair follicle growth. This occurs via dihydrotestosterone (DHT), which inhibits key molecular pathways such as Wnt/β-catenin and Sonic Hedgehog (SHH) signaling. Exosomes derived from plant callus cultures are promising biomaterials for targeted delivery and regenerative medicine. This study aimed to investigate the protective effects of hemp seed callus-derived exosomes (E40) against DHT-induced inhibition of feather follicle development in a chicken embryo model. E40 exosomes were isolated and purified from the calli of germinated hemp seeds. A DHT-induced feather loss model was established by injecting chicken embryos on embryonic day 7 (E7) with DHT (50 ng/mL), with or without co-administration of E40 (40 µg/mL). On embryonic day 12 (E12), feather length, density, and expression of molecular markers were analyzed. The methods included FISH, Western blotting, and quantitative analysis of PTCH1, AR, SHH, SMO, GLI1, Wnt, β-catenin, BMP4, and Noggin. DHT treatment significantly reduced feather length and density. It also downregulated SHH and Wnt/β-catenin markers, upregulating BMP4 and androgen receptor expression. Co-treatment with E40 restored feather length and density to levels comparable to controls and significantly recovered the expression of SHH, SMO, GLI1, Wnt, and β-catenin. E40 also suppressed DHT-induced BMP4 upregulation by approximately 30% and reduced androgen receptor expression. These results suggest that hemp seed-derived exosomes (E40) effectively mitigate DHT-induced feather follicle inhibition by modulating critical signaling pathways and immune-related markers, supporting their potential application as a nanocarrier-based therapeutic strategy for alopecia management.
雄激素性脱发会抑制毛囊生长。这是通过二氢睾酮（DHT）实现的，它会抑制关键分子途径，如Wnt/β-连环蛋白和音猬因子（SHH）信号传导。源自植物愈伤组织培养物的外泌体是用于靶向递送和再生医学的有前景的生物材料。本研究旨在研究大麻籽愈伤组织来源的外泌体（E40）在鸡胚模型中对DHT诱导的毛囊发育抑制的保护作用。从发芽大麻籽的愈伤组织中分离并纯化E40外泌体。通过在胚胎第7天（E7）向鸡胚注射DHT（50 ng/mL），无论是否同时给予E40（40 µg/mL），建立DHT诱导的羽毛脱落模型。在胚胎第12天（E12），分析羽毛长度、密度和分子标志物的表达。方法包括荧光原位杂交（FISH）、蛋白质印迹法以及对PTCH1、AR、SHH、SMO、GLI1、Wnt、β-连环蛋白、BMP4和Noggin的定量分析。DHT处理显著降低了羽毛长度和密度。它还下调了SHH和Wnt/β-连环蛋白标志物，上调了BMP4和雄激素受体表达。与E40共同处理可将羽毛长度和密度恢复到与对照组相当的水平，并显著恢复SHH、SMO、GLI1、Wnt和β-连环蛋白的表达。E40还将DHT诱导的BMP4上调抑制了约30%，并降低了雄激素受体表达。这些结果表明，大麻籽来源的外泌体（E40）通过调节关键信号通路和免疫相关标志物，有效减轻了DHT诱导的毛囊抑制，支持它们作为基于纳米载体的脱发治疗策略的潜在应用。
