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Wnt/β-连环蛋白和ERK信号通路激活:发光二极管光生物调节疗法调控人外根鞘细胞增殖的一种可能机制。

Wnt/β-catenin and ERK pathway activation: A possible mechanism of photobiomodulation therapy with light-emitting diodes that regulate the proliferation of human outer root sheath cells.

作者信息

Kim Jung E, Woo Young J, Sohn Ki M, Jeong Kwan H, Kang Hoon

机构信息

Department of Dermatology, St. Paul's Hospital, College of Medicine, The Catholic University of Korea, 180, Wangsan-ro, Dongdaemun-gu, Seoul, Korea.

出版信息

Lasers Surg Med. 2017 Dec;49(10):940-947. doi: 10.1002/lsm.22736. Epub 2017 Sep 25.

Abstract

BACKGROUND

Outer root sheath cells (ORSCs) play important roles in maintaining hair follicle structure and provide support for the bulge area. The hair growth promoting effects of photobiomodulation therapy (PBMT) have been reported, but the mechanisms for this in human ORCs (hORSCs) have rarely been studied.

OBJECTIVE

The aim of this study was to investigate the effect of various wavelengths of light-emitting diode (LED) irradiation on human ORSCs (hORSCs).

METHODS

LED irradiation effects on hORSC proliferation and migration were examined with MTT assay, BrdU incorporation assay and migration assays. hORSCs were irradiated using four LED wavelengths (415, 525, 660, and 830 nm) with different low energy levels. LED irradiation effects on the expression of molecules associated with the Wnt/β-catenin signaling and ERK pathway, hair stem cell markers, and various growth factors and cytokines in hORSCs were examined with real-time PCR and Western blot assay. The effect of the LED-irradiated hORSCs on cell proliferation of human dermal papilla cells (hDPCs) was examined with co-culture and MTT assay.

RESULTS

PBMT with LED light variably promoted hORSC proliferation and suppressed cell apoptosis depending on energy level. LED irradiation induced Wnt5a, Axin2, and Lef1 mRNA expression and β-catenin protein expression in hORSCs. Phosphorylation of ERK, c-Jun, and p38 in hORSCs was observed after LED light irradiation, and ERK inhibitor treatment before irradiation reduced ERK and c-Jun phosphorylation. Red light-treated hORSCs showed substantial increase in IL-6, IL-8, TNF-a, IGF-1, TGF-β1, and VEGF mRNA. Light irradiation at 660 and 830 nm projected onto hORSCs accelerated in vitro migration. LED-irradiated hORSCs increased hDPCs proliferation when they were co-cultured. The conditioned medium from LED-irradiated hORSCs was sufficient to stimulate hDPCs proliferation.

CONCLUSION

These results demonstrate that LED light irradiation induced hORSC proliferation and migration and inhibited apoptosis in vitro. The growth-promoting effects of LEDs on hORSCs appear to be associated with direct stimulation of the Wnt5a/β-catenin and ERK signaling pathway. Lasers Surg. Med. 49:940-947, 2017. © 2017 Wiley Periodicals, Inc.

摘要

背景

外根鞘细胞(ORSCs)在维持毛囊结构中发挥重要作用,并为毛囊隆突区提供支持。光生物调节疗法(PBMT)促进头发生长的作用已有报道,但在人外根鞘细胞(hORSCs)中的作用机制鲜有研究。

目的

本研究旨在探讨不同波长发光二极管(LED)照射对人外根鞘细胞(hORSCs)的影响。

方法

采用MTT法、BrdU掺入法和迁移实验检测LED照射对hORSC增殖和迁移的影响。使用四种不同低能量水平的LED波长(415、525、660和830nm)照射hORSCs。通过实时PCR和蛋白质免疫印迹法检测LED照射对hORSCs中与Wnt/β-连环蛋白信号通路和ERK通路相关分子、毛囊干细胞标志物以及各种生长因子和细胞因子表达的影响。通过共培养和MTT法检测LED照射的hORSCs对人真皮乳头细胞(hDPCs)细胞增殖的影响。

结果

根据能量水平,LED光的PBMT不同程度地促进hORSC增殖并抑制细胞凋亡。LED照射诱导hORSCs中Wnt5a mRNA、Axin2 mRNA、Lef1 mRNA表达以及β-连环蛋白蛋白表达。LED光照射后观察到hORSCs中ERK、c-Jun和p38磷酸化,照射前用ERK抑制剂处理可降低ERK和c-Jun磷酸化。红光处理的hORSCs中IL-6、IL-8、TNF-α、IGF-1、TGF-β1和VEGF mRNA显著增加。660和830nm的光照射hORSCs可加速其体外迁移。LED照射的hORSCs与hDPCs共培养时可增加hDPCs增殖。LED照射的hORSCs的条件培养基足以刺激hDPCs增殖。

结论

这些结果表明,LED光照射可诱导hORSC增殖和迁移并在体外抑制其凋亡。LED对hORSCs的促生长作用似乎与直接刺激Wnt5a/β-连环蛋白和ERK信号通路有关。《激光外科与医学》49:940 - 947, 2017。© 2017威利期刊公司

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