Light-emitting diode 585 nm photomodulation inhibiting melanin synthesis and inducing autophagy in human melanocytes
Light-emitting diode 585 nm photomodulation inhibiting melanin synthesis and inducing autophagy in human melanocytes
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发光二极管585 nm光调制抑制黑色素合成并诱导人黑素细胞自噬
DOI：
10.1016/j.jdermsci.2017.10.001
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发表时间：
2018-01-01
影响因子：
4.6
通讯作者：
Xiang, Leihong
中科院分区：
文献类型：
--
作者：
Chen, Li;Xu, Zhongyi;Xiang, Leihong
背景：黄褐斑是面部常见的色素沉着皮肤病。发光二极管(LED)光调制(585 Nm)被报道为治疗黄褐斑的有效方法。目的：探讨LED光调制(585 Nm)对人表皮黑素细胞(HEMS)黑素合成的影响。方法：分别用0、5、10和20J/cm(2)585 nm LED照射HEM。处理5d后，用CCK-8法检测细胞存活率，Annexin V APC法检测细胞凋亡率。用分光光度计法测定黑素含量和酪氨酸酶活性。在透射电子显微镜下观察黑素小体分期和自噬小体的变化。共聚焦显微镜下观察到自噬斑点状结构的形成。结果：黄光LED 585 nm对HEMS细胞存活率和细胞凋亡率无明显影响。在5J/cm(2)~20J/cm(2)范围内，LED 585 nm照射可抑制黑素小体成熟，降低黑素含量和酪氨酸酶活性。抑制伴随着酪氨酸酶(TYR)、酪氨酸酶相关蛋白-1(TRP-1)和小眼球相关转录因子(MITF)在mRNA和蛋白水平的表达减少。透射电子显微镜下观察到自噬小体。用发光二极管585 nm光诱导微管相关蛋白轻链3蛋白的自噬斑点结构。LED 585 nm照射后，观察到Lc3构型由Lc3-I转变为Lc3-II，以及p62蛋白的降解。此外，我们还发现LED 585 nm光调制的抗黑素作用可被3-甲基腺嘌呤(3-MA)逆转，3-甲基腺嘌呤(3-MA)通过抑制Ill磷脂酰肌醇3-激酶(PI-3K)抑制自噬形成而抑制自噬。结论：585 nm波长的LED光调制抑制HIMS中的黑色素含量，这种作用是通过抑制黑素合成和诱导HIMS自噬而产生的。这可能为LED光调制在治疗色素沉着障碍中的疗效提供新的见解。(C)2017年，爱思唯尔爱尔兰有限公司代表日本皮肤病研究学会出版。
Background: Melasma is a common hyperpigmentation skin disease on face. Light-emitting diode (LED) photomodulation (585 nm) is reported to be effective for the treatment of melasma. However, whether and how LED photomodulation would influence melanogenesis of human epidermal melanocytes (HEMS) is unknown.Objective: To evaluate the effects of LED photomodulation (585 nm) on melanogenesis in HEMS.Methods: HEMs were irradiated with fluences of 0, 5, 10 and 20 J/cm(2) 585 nm LED light. After 5-day treatment, cell viability was analyzed by CCK-8 assay, and apoptosis was assessed by Annexin V APC assay. Melanin content and tyrosinase activity were measured by spectrophotometer. Melanosome stage and autophagosomes were determined under transmission electron microscope (TEM). The formation of autophagic punctate structures was observed under confocal microscope. RT-PCR and western blotting were used to assess the expression of relative mRNA and protein levels.Results: Yellow light LED 585 nm had no effects on HEMs cell viability and apoptosis. Treatment with LED 585 nm from 5 J/cm(2) to 20 J/cm(2) inhibited melanosome maturation, decreased melanin content and tyrosinase activity. Inhibition was accompanied by the decreased expression of tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1) and microphthalmia-associated transcription factor (MITF) on both mRNA and protein levels. Autophagosomes were observed under TEM. Autophagic punctate structures of microtubule-associated protein light chain 3 (LC3) proteins were induced by LED 585 nm light. The configuration change of LC3 from LC3-I to LC3-II, and the degradation of p62 protein were observed after LED 585 nm. Furthermore, we also revealed that the anti-melanogenic effect of LED 585 nm photomodulation was reversed by 3-Methyladenine (3-MA), which inhibits autophagy by blocking autophagosome formation via the inhibition of type Ill Phosphatidylinositol 3-kinases (PI-3K).Conclusions: Our finding demonstrated that LED photomodulation with 585 nm wavelength suppressed melanin content in HEMs, and the effect was caused by its dose-dependent inhibition on melanogenesis and the induction of HEMs autophagy. This may provide new insights into the efficacy of LED photomodulation in the treatment of hyperpigmentation disorders. (C) 2017 Published by Elsevier Ireland Ltd on behalf of Japanese Society for Investigative Dermatology.