视蛋白3调控窄谱中波紫外线诱导皮肤黑素细胞迁移的作用及机制

窄谱中波紫外线(Narrow-band ultraviolet B,NB-UVB)是目前白癜风的一线治疗方案之一，通过诱导黑素细胞增殖、分化和迁移实现皮损复色，但其确切机制尚不完全明确。近期研究发现，视蛋白3（OPN3）是绿光诱导人眼眶脂肪干细胞迁移的关键分子。但OPN3是否感受NB-UVB并介导表皮黑素细胞的迁移目前未见相关报道。本课题以此为切入点，将以正常人表皮原代黑素细胞为研究对象，在细胞水平研究OPN3的表达与细胞迁移的关系；分别构建过表达和沉默OPN3的慢病毒载体，转染入人原代黑素细胞系中，从分子、蛋白水平研究OPN3对黑素细胞迁移的影响及相关通路；同时体外构建人工组织皮肤模型，利用分子生物学、细胞生物学、组织工程及光化学技术，从组织层面研究OPN3调控人表皮黑素细胞迁移的作用及机制，进一步促进对NB-UVB诱导黑素细胞迁移机制的认识。

Narrow-band UVB (NB-UVB) irradiation has been considered to be one of the most effective treatment options for vitiligo. However, the molecular mechanisms of proliferation, migration and melanogenesis of melanocytes induced by NB-UVB irradiation remain unravelled. Recent studies have shown that opsin3 (OPN3) serve as the photoreceptor of green LED irradiation for the activation of ERK/MAPK/p38 signaling pathway during human orbital fat stem cells migration. However, whether OPN3 senses NB-UVB and mediates the migration of epidermal melanocytes has not been reported. In this research primary human epidermal melanocytes line（HEM-M), which derived from foreskin of children, will be transfected with recombinant lentivirus vectors containing OPN3 gene and OPN3 miRNA respectively. Human tissue-engineered skin will also be exploited for these studies. Gene regulation targeting OPN3 genes is performed with NB-UVB exposure in those cell and mimic skin models. Real-time PCR, Western Blot, flow cytometry and Immunofluorescence were used to analyse the role of OPN3 gene on the migration following NB-UVB irradiation. This study will provide a better understanding of melanocyte migration induced by NB-UVB irradiation in vitiligo.