收稿日期: 2024-02-25
网络出版日期: 2024-11-05
基金资助
国家中医药管理局科技司共建科技项目(GZY-KJS-SD-2023-022);山东省中医药科技重点项目(2021Z021)
The effect of umbilical cord stem cell exosomes on the proliferation of dermal papilla cells
Received date: 2024-02-25
Online published: 2024-11-05
目的 探究人脐带间充质干细胞外泌体（hUCMSC-Exos）对人毛乳头细胞（HDPCs）增殖的影响，并对hUCMSC-Exos促毛发生长作用的机制进行初步探索。 方法 二步酶法分离HDPCs并进行体外培养，培养人脐带间充质干细胞（hUC-MSCs），收集细胞培养上清，通过超高速离心法提取外泌体，并对其进行电镜、粒径及表面标记物鉴定。双氢睾酮（dihydrotestosterone，DHT）诱导HDPCs建立雄激素性脱发细胞模型。将hUCMSC-Exos与HDPCs共培养，采用细胞增殖试验（EdU）检测诱导的HDPCs的相对活力。Real-time qPCR检测碱性磷酸酶（ALP）表达水平、Western blot检测β-catenin、Wnt10b、GSK-3β在蛋白水平的表达。 结果 所获取的HDPCs、hUC-MSCs和hUCMSC-Exos 均符合毛乳头细胞、间充质干细胞及外泌体特征。EdU阳性细胞数显著增加，外泌体可有效促进HDPCs增殖（P < 0.05），增强HDPCs活力，减轻DHT造成的损伤（P < 0.05）。Real-time qPCR显示外泌体可增强ALP基因表达水平（P < 0.05），增强毛囊诱导能力；Western blot证实β-catenin、Wnt10b、GSK-3β在蛋白水平的表达均有差异（P < 0.05）。 结论 hUCMSC-Exos可促进DHT诱导的HDPCs增殖，增强其毛囊再生修复能力，其机制可能与激活Wnt/β-catenin信号通路有关。
罗青 , 黄金金 , 任婷婷 , 周瑞华 , 徐栋花 , 王振华 , 王国颖 . 人脐带干细胞外泌体对人毛乳头细胞增殖的影响[J]. 实用医学杂志, 2024 , 40(20) : 2828 -2834 . DOI: 10.3969/j.issn.1006-5725.2024.20.003
Objective We tried to investigate the effects of human umbilical cord mesenchymal stem cell exosomes （hUCMSC-Exos） on the proliferation of human dermal papilla cells （HDPCs） and the mechanism of hUCMSC-Exos promoting hair growth. Methods HDPCs were isolated using two-step enzymatic method and cultured in vitro. Human umbilical cord mesenchymal stem cells （hUC-MSCs） were cultured. Cell culture supernatant was collected， and exosomes were isolated and extracted using high-speed centrifugation. Electron microscopy， particle size， and surface marker identification were performed on them. Dihydrotestosterone （DHT） induces HDPCs and establishment of an androgenic alopecia cell model. Co-culture hUCMSC-Exos with HDPCs， cell proliferation experiment （EdU） was used to detect the relative activity of induced HDPCs. Real-time qPCR was used to detect the expression level of alkaline phosphatase （ALP）， and Western blot was used to detect β-catenin， Wnt10b， GSK-3β expression at the protein level. Results The obtained primary HDPCs， hUC-MSCs， and hUCMSC-Exos were all conformed to the characteristics of dermal papilla cells， mesenchymal stem cells， and exosomes. The number of EdU positive cells significantly increased， and exosomes could effectively promote the proliferation of HDPCs（P < 0.05）， enhance the vitality of HDPCs and alleviate the damage caused by DHT（P < 0.05）. Real-time qPCR showed that exosomes could enhance the expression level of ALP gene （P < 0.05） and hair follicle induction ability. Western Blot confirmation β-catenin， Wnt10b， GSK-3β were differences in expression at the protein level （P < 0.05）. Conclusions HUCMSC-Exos could promote DHT induced proliferation of HDPCs， enhance their hair follicle regeneration and repair ability， and its mechanism may be related to the activation of Wnt/ β-catenin signaling pathway.
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