Wang Guiyue, Wang Zhili, Zhang Jiaqi, Shen Yan, Hou Xin, Su Lin, Chen Wen, Chen Jiahao, Guo Xiang, Song Hong
School of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, China.
J Cosmet Dermatol. 2023 Nov;22(11):2996-3007. doi: 10.1111/jocd.15890. Epub 2023 Aug 8.
Androgenetic alopecia (AGA) is the most common cause of chronic progressive hair loss in men, and AGA has a severe negative impact on the quality of life and physical and mental health of patients.
Four female C57BL/6 mice were isolated from DP cells in culture (≤4 generations) after stimulation of DPC proliferation by herbal concentrations obtained by the CCK-8 method, and exosomes were isolated by differential centrifugation at low temperature. Testosterone propionate and topical hair removal treatments were used together to establish the C57BL/6 mouse AGA model, which was treated with LTF, 5% minoxidil, and LTF-DPC-EXO, respectively. ELISA was used to detect serum hormone levels, in vivo tracing was used to observe dynamic changes in exosomes, H&E staining showed changes in mouse hair follicle tissue, and (q) RT-PCR and WB were used to detect dorsal skin VEGF, AKT1, and CASP3 expression in dorsal skin tissues.
Hair regeneration was significant in the LTF group, minoxidil group, and LTF-DPC-EXO group mice, and the hair growth was only seen in the local skin in the model group. The hormone T in all treatment groups was lower than that in the model group, and e2 was higher than that in the model group. (q) RT-PCR and western blot showed that VEGF and AKT1 expressions were upregulated and Caspase3 expression was downregulated in the skin sections of mice in the treatment groups.
DPC-EXO obtained through LTF may activate AKT1 and VEGF in the PI3K/AKT signaling pathway to inhibit CASP3, thereby protecting DPC to restore the hair growth.
雄激素性脱发(AGA)是男性慢性进行性脱发的最常见原因,对患者的生活质量和身心健康有严重负面影响。
采用CCK-8法获得草药浓度刺激毛囊乳头细胞(DPC)增殖后,从培养的DPC中分离出4只雌性C57BL/6小鼠(传代≤4代),通过低温差速离心法分离外泌体。联合丙酸睾酮和局部脱毛处理建立C57BL/6小鼠AGA模型,分别用LTF、5%米诺地尔和LTF-DPC-EXO进行处理。采用ELISA检测血清激素水平,采用体内示踪观察外泌体的动态变化,采用苏木精-伊红(H&E)染色观察小鼠毛囊组织变化,采用(定量)逆转录聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法(WB)检测背部皮肤组织中血管内皮生长因子(VEGF)、蛋白激酶B1(AKT1)和半胱天冬酶3(CASP3)的表达。
LTF组、米诺地尔组和LTF-DPC-EXO组小鼠毛发再生明显,模型组仅局部皮肤有毛发生长。各治疗组的激素睾酮(T)均低于模型组,雌二醇(e2)均高于模型组。qRT-PCR和蛋白质免疫印迹结果显示,各治疗组小鼠皮肤切片中VEGF和AKT1表达上调,Caspase3表达下调。
通过LTF获得的DPC-EXO可能激活PI3K/AKT信号通路中的AKT1和VEGF,抑制CASP3,从而保护DPC以恢复毛发生长。