超滤-高效液相色谱联用筛选愈白丸中的抗白癜风活性成分  

Screening of Anti-Vitiligo Active Ingredients in Yubai Pill by Ultrafiltration-High Performance Liquid Chromatography

作　　者：陈龙 吴丛婷 周明[2] 卞芳 李静 CHEN Long;WU Congting;ZHOU Ming;BIAN Fang;LI Jing(Key Laboratory of Featured Preparations of Vitiligo,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang 441021;Key Laboratory of Plant Germplasm Innovation and Specialty Agriculture,Chinese Academy of Sciences,Wuhan 430074)

机构地区：[1]湖北文理学院附属医院,襄阳市中心医院,白癜风特色制剂襄阳市重点实验室,湖北襄阳441021 [2]中国科学院植物种质创新与特色农业重点实验室,湖北武汉430074

出　　处：《分析科学学报》2024年第5期517-523,共7页Journal of Analytical Science

基　　金：湖北省卫健委中医药科研项目(ZY2021Q025);襄阳市中心医院人才启动项目(RC202002)。

摘　　要：愈白丸为本院自制的白癜风口服特效制剂,君药为蒺藜,但有效成分及抗白癜风作用机制缺乏深入研究。本研究分别以酪氨酸酶(TYR)和过氧化氢酶(CAT)为靶点,通过超滤-高效液相色谱法筛选其中的抗白癜风活性成分并探索其作用机制。筛选结果显示,皂苷类物质与TYR亲和力较强,黄酮类物质与CAT亲和力较强,体外酶活性测试证实皂苷类物质能提高TYR的活性,而黄酮类物质则对CAT活力影响较大,其中蒺藜苷元和槲皮素效果最为显著,亲和结合常数分别为0.3846和0.5895,激活率分别为28.24%和43.78%。细胞实验结果显示,当浓度超过5μg/mL时,蒺藜苷元和槲皮素均能促进黑色素细胞增殖,前者可通过激活CAT而降低体内活性氧等自由基水平,后者可激活TYR促进黑色素的合成,两者可在愈白丸治疗白癜风的过程中发挥协同作用。Yubai pill is a homemade featured oral preparation for vitiligo with Tribulus terrestris L.as the monarch drug,but there is a lack of indepth research on the active ingredients and the mechanism of antivitiligo.In this study,ultrafiltrationhigh performance liquid chromatography method was adopted to screen the antivitiligo active ingredients,with tyrosinase(TYR)and catalase(CAT)as the targets respectively,and the mechanisms involved were elucidated.The screen results indicated that saponins had better affinity for TYR and flavonoids preferred CAT.Enzyme activity tests in vitro showed that saponins could enhance the activity of TYR,while flavonoids had a greater effect on CAT activity,with(25R)spirost4ene3,12dione and quercetin possessing the most significant effects,with affinity binding constants of 0.3846 and 0.5895,activation rates of 28.24%and 43.78%,respectively.Cellular experiments showed that when the concentration exceeded 5μg/mL,both(25R)spirost4ene3,12dione and quercetin could promote the proliferation of melanocytes.The former could reduce the level of free radicals such as reactive oxygen species in the body through the activation of CAT,and the latter could activate the TYR to promote the synthesis of melanin,which could play a synergistic role in the treatment of vitiligo with Yubai pill.

关 键 词：白癜风 蒺藜 愈白丸 超滤 亲和筛选 槲皮素 蒺藜苷元 

分 类 号：O657.72[理学—分析化学] R284[理学—化学]