基于细胞水平评价白及多糖对黑色素生成的抑制作用  

The inhibitory effect of Bletilla striata polysaccharide on melanin production based on cellular level

作　　者：蒋勇炫 何毅帆 李征 韩盈盈 刘路阳 李晶洁 Jiang Yongxuan;He Yifan;Li Zheng;Han Yingying;Liu Luyang;Li Jingjie(Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi,Guizhou 563000;College of Pharmacy,Zunyi Medical University,Zunyi,Guizhou 563000;Key Laboratory of Oral Disease Research,Affiliated Stomatology Hospital,School of Stomatology,Zunyi Medical University,Zunyi,Guizhou 563000;Guizhou Yikoubai Technology Co.,Ltd,Guiyang,Guizhou 550081)

机构地区：[1]遵义医科大学,基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563000 [2]遵义医科大学药学院,贵州遵义563000 [3]遵义医科大学口腔疾病研究重点实验室,贵州遵义563000 [4]贵州易口白科技有限公司,贵州贵阳550081

出　　处：《遵义医科大学学报》2023年第10期928-933,共6页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:82060791);贵州省科技计划项目[NO:黔科合支撑(2020)4Y066];遵义医科大学大学生创新创业项目(NO:ZYDC2021009)。

摘　　要：目的研究白及多糖对小鼠黑色素瘤细胞B16-F10中黑色素生成的抑制作用及其机制。方法采用CCK-8法检测白及多糖对B16-F10细胞活性的影响;采用NaOH裂解法测定白及多糖对B16-F10细胞内黑色素含量的影响;采用多巴氧化法和蛋白免疫印迹法(Western blot)分别检测B16-F10细胞中酪氨酸酶(TYR)活性和酪氨酸酶相关蛋白1(TRP-1)、酪氨酸酶相关蛋白2(TRP-2)的表达情况,并利用分光光度仪检测白及多糖的抗氧化能力。结果白及多糖在0.1~1.0 mg/mL浓度内对B16-F10细胞活性无影响。用0.5和1.0 mg/mL白及多糖处理B16-F10细胞,可降低B16-F10细胞内黑色素含量。1.0 mg/mL白及多糖可显著抑制α-MSH诱导的B16-F10细胞中TYR活性;0.5和1.0 mg/mL白及多糖可显著抑制α-MSH诱导的B16-F10细胞中TRP-1、TRP-2的蛋白表达。白及多糖在0.1~1.0 mg/mL浓度内,DPPH平均清除率为18.85%、27.10%、47.92%、60.65%,抗超氧阴离子平均活力为20.94、22.41、30.68、38.86 U/gprot,羟基自由基平均抑制能力为7.39、8.07、18.84、34.37 U/mgprot。结论白及多糖对B16-F10细胞中黑色素的生成具有良好的抑制作用,其机制可能与降低细胞内黑色素含量、抑制TYR活性和TRP-1、TRP-2的蛋白表达以及抗氧化作用有关。Objective This research aimed to investigate the melanin production inhibition and its mechanism of Bletilla striata polysaccharide on melanoma cells(B16-F10).Methods CCK-8 was used to detect the effect of Bletilla striata polysaccharide on the activity of B16-F10 cells.NaOH cracking method was used to determine the effect of Bletilla striata polysaccharide on melanin content in B16-F10 cells.Dopa oxidation method and Western blot were used to determine the effects of Bletilla striata polysaccharide onα-MSH induced the activity of tyrosinase(TYR)and the expression levels of tyrosinase related protein 1(TRP-1)and tyrosinase related protein 2(TRP-2)in B16-F10 cells,respectively.The antioxidant properties of Bletilla striata polysaccharide were detected by spectrophotometry.Results The results showed that Bletilla striata polysaccharide did not affected the ac tivity of B16-F10 cells with the concentration range of 0.1~1.0 mg/mL.B16-F10 cells treated with 0.5 and 1.0 mg/mL Bletilla striata polysaccharide for 24 hours could reduce the content of intracellular melanin.1.0 mg/mL Bletilla striata polysaccharide could significantly inhibit the activity of TYR thatα-MSH induced in B16-F10 cells.And the 0.5 and 1.0 mg/mL Bletilla striata polysaccharide could significantly inhibit the expression levels of TRP-1,TRP-2 thatα-MSH induced in B16-F10 cells.The average clearance rate of DPPH are 18.85%,27.10%,47.92%,60.65%,the average activity of anti superoxide anion are 20.94,22.41,30.68,38.86 U/gprot,the average inhibition capacity of hydroxyl radical are 7.39,8.07,18.84,34.37 U/mgprot with the concentration range of 0.1~1.0 mg/mL.Conclusion Bletilla striata polysaccharide had a good inhibitory effect on melanin production,and its mechanism may be related to the content reduction of intracellular melanin,the inhibition of TYR activity,the decrease of the TRP-1 and TRP-2 expression levels and antioxidation capacity.

关 键 词：白及多糖 黑色素 小鼠黑色素瘤细胞 酪氨酸酶 抗氧化 

分 类 号：R965.1[医药卫生—药理学]