视蛋白3调控长波紫外线诱导皮肤黑素生成的作用及机制

长波紫外线（UVA）是调控皮肤黑素生成的重要外源性环境因素，但人表皮黑素细胞是如何感受UVA光信号并转化为细胞内色素合成信号的机制，迄今尚不完全清楚。近期研究表明，黑素细胞是视蛋白（OPN）介导的人表皮紫外线光传导受体细胞，本课题组前期研究证实了人表皮黑素细胞OPN3的表达高低与酪氨酸酶的活性呈现相关性。本课题以此为切入点，构建含OPN3的重组慢病毒载体和含OPN3的沉默RNA，转染正常人表皮黑素细胞，同时构建含病毒转染的正常人表皮黑素细胞的组织工程皮肤和皮肤外植体，并用UVA干预对OPN3的表达进行调控，采用real-time PCR、Western Blot、流式细胞术、免疫荧光、质谱分析，研究OPN3基因调控对黑素细胞UVA色素合成的影响及褐黑素、真黑素含量的调节，阐明OPN3调控UVA诱导人表皮黑素细胞黑素生成的作用及机制，为色素障碍性疾病及皮肤光老化的研究提供实验和理论基础。

Long-wave ultraviolet (UVA) is the most important exogenous environmental factor regulating the melanogenesis. However, how human epidermal melanocytes sense UVA light signals and convert them into intracellular pigment synthesis signals is still not fully understood. Recent studies have shown that melanocytes are photoreceptor cells medicated by OPNs in human skin. Our previous study confirmed that the expression of OPN 3 in human epidermal melanocytes is correlated with the activity of tyrosinase. Primary human epidermal melanocytes,which derived from foreskin of children, will be transfected with recombinant lentivirus vectors containing OPN 3 gene and OPN 3 miRNA respectively. Human skin explant and human tissue-engineered skin will also be exploited for these studies. Gene regulation targeting OPN 3 genes is performed with UVA exposure in those cell and mimic skin models. Real-time PCR, Western Blot, flow cytometry, Immunofluorescence and mass spectrometry were used to analyse the role of OPN 3 gene on the melanogenesis following UVA irradiation. This study will help for the understanding of the mechanism of UVA pigment synthesis in the skin. And it will provide experimental and theoretical basis for future study of pigmentation disorders and skin photoaging.