摘要
目的研究烟酰胺作用于人皮肤黑素细胞后,对黑素细胞中钙信号、细胞骨架的影响及其与黑素小体运动速率和分布的作用,并探讨其中的相互关系。方法体外培养的人皮肤黑素细胞中分别加入0.00、0.05、0.25、1.25、6.25mg/ml烟酰胺后,用荧光抗体染色法检测黑素细胞中游离钙离子浓度,以生化法测定细胞中钙泵活性,以Western Blot蛋白浓度测定法测定动力蛋白Dynein的表达水平,显微镜下观察活细胞中烟酰胺对黑素小体运动速率及黑素小体分布的影响。结果烟酰胺对人皮肤黑素细胞中游离钙浓度影响显著,并有剂量-效应关系,y=76.4612-5.435x(r=0.97);细胞钙泵活性随烟酰胺浓度的增加而下降;细胞中动力蛋白表达随烟酰胺加入的浓度不同而改变;在烟酰胺浓度为0.05和25mg/ml时,细胞功能正常的情况下,钙泵活性正常,能够调控细胞中的物质转运,而黑素细胞中Dynein表达却明显减少(P<0.05),而烟酰胺浓度为1.25mg/ml时,钙泵活性减弱(P<0.05),黑素细胞中动力蛋白Dynein的表达增加(P<0.05)。当烟酰胺作用浓度为0.25和1.25mg/ml时,黑素细胞中黑素小体的运动速率明显增加(P<0.05),同时细胞中黑素小体密集度随烟酰胺浓度增加而下降。结论烟酰胺参与黑素的转运过程并能加快黑素细胞中黑素小体的运动速率,显示烟酰胺对钙泵、细胞动力蛋白Dynein表达和黑素小体运动具有调控作用,作用浓度为一重要调节因素。
Objective To study the effects of nicotinamide on the human skin melanocytes and try to explore the potential mechanism of nicotinamide on the calcium signal transduction, cytoskeleton. Methods The primary cultured human skin melanocytes from foreskin were selected as the target cells in the present study. 0.00, 0.05, 0.25, 1.25 and 6.25 mg/ml nicotinamide were applied respectively. Western blot, fluorescent probes （Flu-3AM）, flow cytometry analysis and time-lampse microscope digital skill were used to evaluate the effects of nicotinamide on melanosome motility and the melanosome distribution in melanocytes. Results The results showed that nicotinamide had a potential effect on regulating free calcium concentration in a dose-dependent manner （y=76.461 2-5.435x, r=0.97）;The activity of Na^＋-K^＋-Ca^2＋-ATPase was down regulated with the increasing concentration of nicotinamide. The expression of cellular dynein was also altered by nicotinamide; Na^＋-K^＋-Ca^2＋-ATPase activity was kept normal when given 0.05, 0.25 mg/ml nicotinamide, while the dynein protein expression was inhibited （P〈0.05）,Na^＋-K^＋-Ca^2＋-ATPase activity was decreased significantly when given 1.25 mg/m] nicotinamide （P〈0.05）, while the dynein protein expression was increased （P 〈0.05）. When treated with 0.25 mg/ml and 1.25 mg/ml concentration of nicotinamide, movement velocity in melanocytes was accelerated and the distribution characters of melanosomes were different among groups. Conclusion Nicotinamide may mediate the melanin transport in melanocytes in vitro. The concentration of nicotinamide plays a key role in the melanin regulating path. Nicotinamide could be used as a regulator for melanin transport.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2009年第8期680-683,共4页
Journal of Environment and Health
基金
新疆维吾尔自治区科技厅资助项目(200623113)
作者简介
帕它木·莫合买提（1972-），女，副教授，从事皮肤生理毒理学研究。
