谷氨酸信号通路对黑素细胞黑素转运的作用  

Roles of glutamate signaling pathway in melanin transfer

作　　者：高丽丽[1] 刘晶 邹伟[3] 刘鹏 张媛[3] 高船舟[4] 王楠[1] 宋智琦[1] 

机构地区：[1]大连医科大学附属第一医院皮肤科,116021 [2]中英再生医学应用研究中心 [3]辽宁师范大学生命科学学院生物技术与分子药物研发重点实验室 [4]大连医科大学中心实验室

出　　处：《中华皮肤科杂志》2012年第10期700-703,共4页Chinese Journal of Dermatology

基　　金：基金项目：国家自然科学基金（30670650,81171491）;辽宁省自然科学基金（20042137）;辽宁省教育厅课题（2008167）;大连市科技局民生科技项目（2010E11SF008）

摘　　要：目的探讨谷氨酸信号通路在黑素转运中的作用。方法原代培养并纯化黑素细胞及角质形成细胞，免疫荧光显微镜观察离子型谷氨酸受体N-甲基-D-天冬氨酸受体1（NMDARl）和N-甲基-D-天冬氨酸受体2A（NMDAR2A）在黑素细胞内的分布，共聚焦显微镜观察100μmo]／LNMDAR激动剂NMDA和100μmol／L拈抗剂地卓西平马来酸盐（dizocilpinemaleate，MK801）作用5min和1h后黑素细胞内钙离子浓度的变化以及100μmol／LMK801对黑素细胞内微管蛋白的影响。结果100μmol／LNMDA可使黑素细胞内瞬时钙离子浓度升高，但100μm01]LMK801可使其降低；MK801先作用于黑素细胞5min或1h阻断NMDA受体后，NMDA均不能再次诱导瞬时钙离子浓度升高。共聚焦显微镜观察发现MKS01作用24h后，胞内微管蛋白重新分布聚集于核周。扫描电镜观察发现100μmol／LMK801作用于黑素细胞一角质形成细胞共培养体系48h后，黑素细胞和角质形成细胞之间以及两种细胞表面的丝状伪足数量明显减少。共培养体系下，100μmol/LMK801作用后，角质形成细胞中的黑素含量明显降低，即从黑素细胞向角质形成细胞转移的黑素数量明显减少。结论谷氨酸信号通路对黑素细胞胞内钙离子浓度、微管蛋白分布、黑素细胞伪足形成以及黑素细胞及角质形成细胞间的黑素转运具有一定调节作用。Objective To investigate the roles of glutamate signaling pathway in melanin transfer. Methods Epidermal melanocytes and keratinocytes were isolated from human foreskin tissue followed by purification and primary culture. Immunofluorescence microscopy was conducted to observe the intracellular distribution of N-methy-D-aspartate receptor 1 （NMDAR1） and NMDAR2A in melanoeytes. Some melanocytes were classified into 4 groups to be pretreated with MKS01 （the NMDAR antagonist dizocilpine maleate） at 100μmol/L for 5 minutes followed by treatment with NMDA （an NMDAR agonist） at 100μmol/L （MK801- pretreated group 1）, pretreated with MKSO1 at 100 μmol/L for 1 hour followed by treatment with NMDA at 100 μmol/L （MK801-pretreated group 2）, treated with MK801 at 100 μ mol/L for 5 minutes （MK801 group）, treated with NMDA at 100 μmol/L for 5 minutes （NMDA group）, respectively, then, confocal microscopy was performed to measure the intracellular calcium （Ca2＋） concentration of the melanocytes. The distribution of 13-tubulin was visualized by confocal microscopy in melanoeytes treated with MK801 at 100 μmol/L for 24 hours. Some melanocytes and keratinocytes were cocultured with or without MK801 at 100μmol/L for 24 or 48 hours, then, scaning microscopy was carried out to observe the junction structure between melanocytes and keratinocytes, and alkali method coupled with spectrophotometric analysis to determine melanin content in keratinoeytes. Results The intracellular calcium concentration of melanocytes was decreased by MK-801, but increased by NMDA at 100 μ mol/L, and the increase was blocked by the pretreatment with MK-801 for 5 minutes or 1 hour. After incubation with MK-801 at 100 μmol/L for 24 hours, a more intense staining for β-tubulin was observed around the nuclei of melanocytes. There was a significant reduction in the number of filopodia on the surface Of and between melanocytes and keratinoeytes after treatment with MK-801 at 100μmol/L for 48 hours. Also, the content of m

关 键 词：受体 谷氨酸 黑素细胞 伪足 黑素类 角蛋白细胞 微管蛋白 

分 类 号：R758.4[医药卫生—皮肤病学与性病学]