TMT-Based Quantitative Proteomic Analysis Reveals the Effect of Bone Marrow Derived Mesenchymal Stem Cell on Hair Follicle Regeneration.
TMT-Based Quantitative Proteomic Analysis Reveals the Effect of Bone Marrow Derived Mesenchymal Stem Cell on Hair Follicle Regeneration.
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DOI：
10.3389/fphar.2021.658040
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发表时间：
2021
影响因子：
5.6
通讯作者：
Xu X
中科院分区：
文献类型：
--
作者：
Zhang C;Li Y;Qin J;Yu C;Ma G;Chen H;Xu X
脱发（HL）是一种常见的慢性问题，其病因尚不明确。在此，我们探讨了骨髓间充质干细胞（BMSC）和条件培养基（MSC-CM）作为毛囊增殖和再生调节剂的功能，以及这种活性的机制基础。通过诱导多谱系分化并使用 CCK-8 试剂盒在体外培养和鉴定 BMSC。然后向小鼠背部皮肤注射 BMSC 和 MSC-CM，然后通过苏木精和伊红 (H&E) 染色和免疫荧光 (IF) 染色评估这些注射对毛周期转变和毛囊干细胞 (HFSC) 增殖的影响。然后，我们对对照小鼠和接受 BMSC 或 MSC-CM 治疗的小鼠进行基于串联质量标签 (TMT) 的定量蛋白质组学分析，以鉴定与这些治疗相关的差异表达蛋白 (DEP)。平行反应监测（PRM）被用作验证我们的蛋白质组分析结果的一种手段。在此，我们发现BMSC和MSC-CM注射导致休止期毛囊向毛发生长初期毛囊的转变，并且我们观察到Krt15和Sox9阳性的HFSC的增殖增强。当比较 BMSC 与对照组以及 MSC-CM 与对照组时，我们的 TMT 分析分别确定了 1,060 和 770 个 DEP（倍数变化＞1.2 或＜0.83 且 p ＜ 0.05）。随后对 14 个选定的 DEP 进行的 PRM 验证证实了这些发现，并确定了 Stmn1、Ncapd2、Krt25 和 Ctps1 作为蛋白质-蛋白质相互作用网络中的枢纽 DEP。总之，这些数据表明 BMSC 和 MSC-CM 治疗可以促进 HFSC 增殖，从而促进毛囊再生。我们的蛋白质组学分析进一步表明，Krt25、Cpm、Stmn1 和 Mb 可能在这种情况下在毛囊转变中发挥核心作用，并且可能代表治疗 HL 的可行临床靶点。
Hair loss (HL) is a common chronic problem of poorly defined etiology. Herein, we explored the functionality of bone marrow-derived mesenchymal stem cell (BMSC) and conditioned medium (MSC-CM) as regulators of hair follicle proliferation and regeneration, and the mechanistic basis for such activity. BMSC were cultured and identified in vitro through the induction of multilineage differentiation and the use of a CCK-8 kit. The dorsal skin of mice was then injected with BMSC and MSC-CM, and the impact of these injections on hair cycle transition and hair follicle stem cell (HFSC) proliferation was then evaluated via hematoxylin and eosin (H&E) staining and immunofluorescent (IF) staining. We then conducted a tandem mass tags (TMT)-based quantitative proteomic analysis of control mice and mice treated with BMSC or MSC-CM to identify differentially expressed proteins (DEPs) associated with these treatments. Parallel reaction monitoring (PRM) was utilized as a means of verifying our proteomic analysis results. Herein, we found that BMSC and MSC-CM injection resulted in the transition of telogen hair follicles to anagen hair follicles, and we observed the enhanced proliferation of HFSCs positive for Krt15 and Sox9. Our TMT analyses identified 1,060 and 770 DEPs (fold change＞1.2 or＜0.83 and p ＜ 0.05) when comparing the BMSC vs. control and MSC-CM vs. control groups, respectively. Subsequent PRM validation of 14 selected DEPs confirmed these findings, and led to the identification of Stmn1, Ncapd2, Krt25, and Ctps1 as hub DEPs in a protein-protein interaction network. Together, these data suggest that BMSC and MSC-CM treatment can promote the proliferation of HFSCs, thereby facilitating hair follicle regeneration. Our proteomics analyses further indicate that Krt25, Cpm, Stmn1, and Mb may play central roles in hair follicle transition in this context and may represent viable clinical targets for the treatment of HL.
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