Influence of endothelin to cell morphology of dendrites and cell motility in cultured normal human melanocytes

黑素细胞在创伤愈合过程中可以增殖和迁移，但其机制尚不清楚，其主要是受周围角质形成细胞产生的内皮素-1（endotherin-1，ET-1）的影响，黑素细胞的合成和树突状化，为角质形成细胞提供黑素以防御紫外线损伤。本研究通过观察内皮素-1（ET-1）对黑素细胞迁移率、树突状及参与局部接触形成的局部接触蛋白p125 FAK表达的影响，采用延时图像分析技术，观察了含ET-1的培养液对黑素细胞迁移率及树突形态的影响。用荧光素和罗丹明双染法检测p125 FAK的表达，用真实的RT-PCR检测p125 FAK mRNA的表达，发现在无ET-1的情况下，黑素细胞的迁移率在细胞接种后7小时最高，细胞迁移率随ET-1浓度的增加而降低。黑素细胞在细胞壁内肌动蛋白应力纤维的周围显示出含有p125 FAK的斑块，然而，p125 FAK的斑块数量被ET-1浓度减少。ET-1还可降低p125 FAK mRNA的表达。而黑素细胞的树突长度、树突百分比和树突数则随ET-1浓度的增加而增加，表明表皮角质形成细胞旁分泌的ET-1在黑素细胞运动和树突中的作用。我们的数据进一步支持了角质形成细胞控制皮肤中黑素细胞增殖或分化的概念，并可用于治疗白癜风，炎症后色素脱失。

Melanocyte can proliferate and migrate in wound healing, however, the mechanism of it remains unknown Synthesis of melanin and dendricity of melanocytes, which supply melanin for keratinocytes to defense against UV damageis, influenced by endotherin-1 (ET-1) that produced by surrunded keratinocytes. Then we study the influence of ET-1 to melanocytes migration, dendricity and expression of p125 FAK, which is known as one of the focal contct localized protein involved in formation of focal contact.Mean migration rates and dendrites morphology of human newborn melanocytes cultured in ET-1 contained medium on a cover-slips coated fibronectin were mesured using the time-lapse image analysis. Expression of p125 FAK by double stained with anti-p125 FAK fluolescein and rhodamine, and of p125 FAK m-RNA by real time RT-PCR.under the ET-1 contained medium were also examined.Highest migration rate of melanocyte under no ET-1 was noted in 7 hrs after cell seeding.Reduced cell migration dependent on concentration of endothelin-1 was exibited. Melanocytes exhibited p125 FAK containing plaques localized in the periphery of actin stress fiber within the cell wall, however, plaque number of p125 FAK was reduced by ET-1 concentration. p125 FAK m-RNA was also reduced by ET-1. The otherhand length of dendrites, %dendricity, and number of dendrites of the melanocytes were increased dependent on ET-1 concentration.Taken together, we demonstrated roles of ET-1, which is paracrined by keratinocytes, in melanocyte motility and dendricity. Our data provide further support for the concept that proliferation or differentiation of melanocyte in the skin is controlled by keratinocyte, and can be usefull for the treatment for the vitiligo vulugalis, postinflammatory depigmentation.