Metformin Promotes the Hair-Inductive Activity of Three-Dimensional Aggregates of Epidermal and Dermal Cells Self-Assembled in vitro
Metformin Promotes the Hair-Inductive Activity of Three-Dimensional Aggregates of Epidermal and Dermal Cells Self-Assembled in vitro
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DOI：
10.1159/000521400
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发表时间：
2021-12
影响因子：
2.7
通讯作者：
Chao Sun;Shuangye Hu;Bing Dong;Shan Jiang;F. Miao;T. Lei
中科院分区：
文献类型：
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作者：
Chao Sun;Shuangye Hu;Bing Dong;Shan Jiang;F. Miao;T. Lei
简介：虽然已有报道抗糖尿病药物二甲双胍具有抗氧化、抗衰老甚至抗肿瘤等多种超降血糖活性，局部应用二甲双胍也可诱导毛发再生，但其确切机制尚不清楚。目的：本研究的目的是评估二甲双胍对小鼠毛囊重建模型中毛发生长的影响，该模型由体外自组装的表皮和真皮细胞（DC）三维聚集体（3D聚集体）生成。方法：从50只1日龄C57 BL/6小鼠皮肤中分离培养表皮细胞和树突状细胞。为了追踪DC在3D聚集体的自组装过程中的分布，用Vybrant Dil Cell-Labeling Solution标记DC并以1：1的比例与表皮细胞混合。用10 mM二甲双胍处理形成的3D聚集体，然后将其移植到受体BALB/c裸鼠中。使用免疫荧光染色、定量实时RT-PCR（qRT-PCR）和蛋白质印迹法，在移植皮肤组织和二甲双胍处理的培养3D聚集体中检测与DC毛发诱导活性相关的生物标志物（肝细胞生长因子[HGF]、β-连环蛋白-1 [CD 133]、碱性磷酸酶[ALP]、β-连环蛋白和SRY盒转录因子2 [SOX 2]）。采用qRT-PCR和Western blotting检测不同代次DC中CD 133的表达水平。结果如下：Metalloy直接刺激培养的3D聚集体的ALP活性，上调分子标记物（HGF、CD 133、ALP、β-catenin和SOX 2）的蛋白和mRNA表达水平，并提高重建毛囊的存活率。此外，我们还发现二甲双胍增加了DC中CD 133的表达，从而维持了它们通常会因连续传代培养而丧失的促凋亡能力。结论：这些结果表明，二甲双胍可以通过上调DC的毛发诱导能力来促进体外毛囊再生，从而进一步评估男性或女性型脱发的临床治疗。
Introduction: Although it has been reported that the antidiabetic drug metformin has multiple extra-hypoglycemic activities, such as anti-oxidation, antiaging, and even antitumor, topical metformin also can induce hair regeneration, but the precise mechanism involved in that process is still unclear. Objectives: The aim of this study was to assess the effect of metformin on hair growth in a mouse hair-follicle reconstitution model generated by in vitro self-assembled three-dimensional aggregates of epidermal and dermal cells (DCs) (3D aggregates). Methods: Epidermal cells and DCs were isolated and cultured from the mouse skin of 50 C57BL/6 mouse pups (1-day-old). For tracing the distribution of DCs during the self-assembly process of 3D aggregates, the DCs were labeled with Vybrant Dil Cell-Labeling Solution and mixed with epidermal cells at a 1:1 ratio. Formed 3D aggregates were treated with 10 mM metformin and then were grafted into recipient BALB/c nude mice. The biomarkers (hepatocyte growth factor [HGF], prominin-1 [CD133], alkaline phosphatase [ALP], β-catenin, and SRY-box transcription factor 2 [SOX2]) associated with the hair-inductive activity of DCs were detected in the grafted skin tissues and in cultured 3D aggregates treated with metformin using immunofluorescent staining, quantitative real-time RT-PCR (qRT-PCR), and Western blotting. Furthermore, the expression levels of CD133 were also examined in DCs with different passage numbers using qRT-PCR and Western blotting. Results: Metformin directly stimulates the activity of ALP of cultured 3D aggregates, upregulates both the protein and mRNA expression levels of molecular markers (HGF, CD133, ALP, β-catenin, and SOX2), and improves the survival rate of reconstituted hair follicles. Moreover, we also found that metformin increases the expression of CD133 in DCs thus maintaining their trichogenic capacity that would normally be lost by serial subculture. Conclusions: These results suggest that metformin can promote hair follicle regeneration in vitro through upregulation of the hair-inductive capability of DCs, warranting further evaluation in the clinical treatment of male or female pattern hair loss.