水解珍珠液抑制人皮肤细胞黑色素形成的作用机制研究  

Study on the Mechanism of Hydrolyzed Pearl Solution Inhibiting Melanin Formation in Human Skin Cells

作　　者：韩丝银 张晶晶 岑妍慧 赖伟辉 杨瑞 林江 Han Siyin;Zhang Jingjing;Cen Yanhui;Lai Weihui;Yang Rui;Lin Jiang(School of Basic Medicine,Guangxi University of Chinese Medicine,Nanning 530200,China;College of Orthopedics and Traumatology,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学基础医学院,广西南宁530200 [2]广西中医药大学骨伤学院,广西南宁530200

出　　处：《亚太传统医药》2023年第12期26-33,共8页Asia-Pacific Traditional Medicine

基　　金：广西高校中青年教师科研基础能力提升项目(2019KY0312);广西高校中青年教师科研基础能力提升项目(2021KY0291);广西中医药大学博士科研启动基金项目(2018BS013);广西中医药大学广西一级学科建设开放项目(2019xk058);广西自然科学基金项目(2020GXNSFAA259011);中医学广西一流学科(桂教科研[2022]1号);“全国高校黄大年式教师团队”-广西中医药大学中西医基础课程教师团队(教师函[2022]2号);广西名中医林江传承工作室(广西中医科教发展[2021]6号)。

摘　　要：目的:明确水解珍珠液抑制人皮肤细胞黑色素形成的作用机制。方法:水解珍珠液培养不经过或经过UVB照射的人角质形成细胞(Hacat)48 h后,CCK8法检测细胞增殖率,生化法检测细胞ROS、MDA含量,ELISA法检测细胞PC、NT含量;水解珍珠液培养不经过或经过α-MSH刺激的人黑色素瘤细胞(A875)48 h后,CCK8法检测细胞增殖率,NaOH法检测细胞内黑色素含量,Western blot法检测细胞TYR及TRP-1蛋白的表达;水解珍珠液培养不经过任何刺激或经过α-MSH刺激或经过UVB照射的A875/Hacat混合细胞48 h后,倒置光学显微镜检测各组A875细胞中树突的形成,倒置荧光显微镜检测A875细胞中TRP-1阳性黑素体转移到Hacat细胞的情况。结果:UVB照射Hacat细胞显著抑制增殖率,而水解珍珠液对Hacat细胞没有毒性,显著降低了UVB照射下Hacat细胞的ROS、MDA、PC、NT含量(P<0.05),提高细胞增殖率(P<0.05);α-MSH刺激显著提高A875细胞的黑色素含量(P<0.05),显著降低A875细胞的增殖率(P<0.05),水解珍珠液培养不经过或经过α-MSH刺激的A875,结果都显著降低细胞TYR和TRP-1蛋白表达及黑色素含量(P<0.05),且随着水解珍珠液浓度提高依赖性降低。水解珍珠液培养不经过任何刺激或经过α-MSH刺激或经过UVB照射的A875/Hacat混合细胞中,A875细胞树突形成水平均降低,A875细胞中TRP-1阳性黑素体转移到Hacat细胞的水平均降低,且随着水解珍珠液浓度提高而依赖性降低。结论:水解珍珠液能够抑制人皮肤细胞黑色素形成,该机制与黑色素合成期间,水解珍珠液抑制TYR和TRP-1蛋白表达,降低黑色素含量有关;同时,与黑色素合成之后,水解珍珠液抑制黑素体从黑素细胞转移到角质细胞也有关。Objective:To clarify the mechanism of hydrolyzed Pearl solution inhibiting melanin formation in human skin cells.Methods:Human keratinocytes(Hacat)without or after UVB irradiation were cultured in Pearl hydrolysate for 48 hours.The cell proliferation rate was detected by CCK8 method,the contents of ROS and MDA were detected by biochemical method,and the contents of PC and NT were detected by ELISA;After human melanoma cells(A875)without or stimulated were cultured for 48 hours byα-MSH in Pearl hydrolysate,the cell proliferation rate was detected by CCK8 method,the content of melanin in cells was detected by NaOH method,and the expression of TYR and TRP-1 protein in cells was detected by Western blot method;After the mixed A875/Hacat cells without any stimulation or stimulated byα-MSH or irradiated by UVB were cultured in Pearl hydrolysate for 48 hours,the formation of dendrites in A875 cells in each group was detected by inverted light microscope,and the transfer of TRP-1 positive melanosomes in A875 cells to Hacat cells was detected by inverted fluorescence microscope.Results:UVB irradiation significantly inhibited the proliferation rate of Hacat cells,while Pearl hydrolysate had no toxicity to Hacat cells.It significantly reduced the contents of ROS,MDA,PC,NT of Hacat cells under UVB irradiation(P<0.05),and increased the cell proliferation rate(P<0.05);Theα-MSH stimulation significantly increased the melanin content of A875 cells(P<0.05)and significantly reduced the proliferation rate of A875 cells(P<0.05).The Pearl hydrolysate culturing A875 cells without or afterα-MSH stimulation significantly reduced the expression of TYR and TRP-1 proteins and melanin content(P<0.05),which decreased dependently with the increase of the concentration of hydrolyzed Pearl solution.In A875/Hacat mixed cells cultured in hydrolyzed Pearl solution without any stimulation or stimulated byα-MSH or irradiated by UVB,the level of dendritic formation in A875 cells decreased,and the level of TRP-1 positive melanosome transferred to

关 键 词：水解珍珠液 A875 HACAT Α-MSH 黑色素 

分 类 号：R285.5[医药卫生—中药学]