天然产物研究与开发 ›› 2015, Vol. 27 ›› Issue (4): 578-584.doi: 10.16333/j.1001-6880.2015.04.005

• 研究论文 • 上一篇    下一篇

制何首乌提取物及主要单体成分对细胞酪氨酸酶及抗氧化保护作用研究

冯光远,石璐缘,崔宝弟,李登科,孙震晓*   

  1. 北京中医药大学中药学院生物制药系,北京 100102
  • 出版日期:2015-04-28 发布日期:2015-05-08

Effect of Water Extract and Main Monomer Components of Processed Polygonum multiflorum on Intracellular Tyrosinase and Their Antioxidant Activity

FENG Guang-yuan,SHI Lu-yuan,CUI Bao-di,LI Deng-ke,SUN Zhen-xiao*   

  1. Department of Biopharmaceutical Sciences,College of Chinese Pharmacy,Beijing University of Chinese Medicine,Beijing 100102,China
  • Online:2015-04-28 Published:2015-05-08

摘要: 为了探究制何首乌发挥乌发功效的机制,我们利用多巴速率氧化法、多巴染色法评价制何首乌水提物(PWE)、粗多糖及二苯乙烯苷、大黄素对B16F10中酪氨酸酶活性的影响,并利用实时荧光定量PCR法检测其对细胞酪氨酸酶mRNA表达量的影响。结果显示,受试药物对B16F10内酪氨酸酶活性无明显激活作用,但500 μg/mL的PWE可显著激活酪氨酸酶mRNA表达;另一方面,利用DPPH自由基清除法测定PWE及大黄素、二苯乙烯苷的体外抗氧化能力,并利用HepG2C8细胞模型考察其在细胞水平上的抗氧化能力,结果发现,PWE及二苯乙烯苷体外可浓度依赖地清除DPPH自由基,且能激活模型细胞HepG2C8中ARE的表达,表明二者具有显著的抗氧化活性,或为制何首乌乌发作用要因。

关键词: 制何首乌水提物, 制何首乌粗多糖, 大黄素, 二苯乙烯苷, 酪氨酸酶, 抗氧化

Abstract: The aim of this study was to explore the mechanism of processed Polygonum multiflorum induced hair blackening.The effects of water extract of processed P.multiflorum (PWE),crude polysaccharide from processed P.multiflorum, 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (THSG) and emodin on tyrosinase activity in B16F10 cells were assayed by measuring the oxidation rate of L-DOPA and Dopa staining testing.The impact of PWE,THSG and emodin on the tyrosinase mRNA expression in B16F10 cells was also examined using qRT-PCR.The results showed that none of the tested substance activated the tyrosinase activity effectively,though PWE (500 μg/mL) stimulated tyrosinase mRNA expression significantly.Then,the antioxidant capacity of PWE,emodin and THSG was determined using DPPH radical scavenging assay and the HepG2C8 cells.It showed that PWE and THSG eliminated DPPH free radical in a dose-dependent manner in vitro and elevated the expression of ARE in HepG2C8 cells.Therefore,the results in this report indicated that the strong antioxidant activity might be the reason for processed P.multiflorum to cause hair blackening.

Key words: water extract of processed Polygonum multiflorum;crude polysaccharide of processed Polygonum multiflorum, emodin;2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside, tyrosinase;antioxidant

中图分类号: 

Q285.5