麻黄非生物碱部位抗过敏性哮喘活性组分的化学成分分析  

Analysis of Chemical Components of Anti-allergic Asthma Active Fractions in Alkaloids-free Part of Ephedrae Herba

作　　者：靳京妹 于大永[1] 侯滔[2] 张秀莉 史丽颖[1] JIN Jingmei;YU Dayong;HOU Tao;ZHANG Xiuli;SHI Liying(School of Life Sciences and Biotechnology,Dalian University,Liaoning Dalian 116622,China;Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Liaoning Dalian 116023,China)

机构地区：[1]大连大学生命科学与技术学院,辽宁大连116622 [2]中国科学院大连化学物理研究所,辽宁大连116023

出　　处：《中国药房》2020年第9期1068-1073,共6页China Pharmacy

基　　金：辽宁省自然科学基金指导计划项目(No.201602027)。

摘　　要：目的:为阐明麻黄非生物碱部位抗过敏性哮喘的药效物质基础提供参考。方法:麻黄用85%乙醇提取、正庚烷萃取后经固相萃取(填料为AC18)前处理,对麻黄提取物进行非生物碱部位富集,并采用高效液相色谱法经Unitary C18柱和Eclipse XDB-C18色谱柱进行麻黄非生物碱类组分制备。以HT-29细胞中高表达的G蛋白偶联受体35(GPR35受体)为靶标、GPR35受体激动剂敏喘宁(1μmol/L)为阳性对照、动态质量重置(DMR)响应值为检测指标,通过无标记细胞整合药理学方法考察各组分(质量浓度均为100μg/m L)对GPR35受体的激动活性和脱敏活性,以筛选抗过敏性哮喘的活性组分。应用高效液相色谱-四极杆飞行时间质谱(HPLC-Q-TOF-MS)技术对活性组分的化学成分进行鉴定。结果:麻黄非生物碱部位主要分为固相萃取前处理的沉淀部分和95%甲醇洗脱部分,从上述两部分中共富集得到了20个组分。其中,沉淀部分中组分F1.5~F1.10及95%甲醇洗脱部分中组分F2.5~F2.10对GPR35受体有较强的激动活性,同时对GPR35受体激动剂敏喘宁显示出了较强的脱敏活性;并且沉淀部分中组分F1.5~F1.10在HT-29细胞上引起的DMR响应信号强度甚至超过了阳性对照药敏喘宁。经HPLC-Q-TOF-MS分析,共从活性组分中鉴定出了24个化合物,其中黄酮类14个、挥发油类2个、有机羧酸类7个、蒽醌类1个。结论:麻黄非生物碱部位主要以黄酮类成分为主,其具有一定的抗过敏性哮喘活性。OBJECTIVE:To provide reference for elucidating the anti-allergic asthma constituents in alkaloids-free part of Ephedrae Herba.METHODS:Ephedrae Herba was extracted with 85%ethanol and n-heptane,and then subjected to solid-phase extraction(filler AC18)for pretreatment to enrich alkaloids-free part from the extract of Ephedrae Herba.HPLC method was adopted,and alkaloids-free fractions of Ephedrae Herba were performed on Unitary C18 column and Eclipse XDB-C18 column.Using high expression G protein coupled-receptor 35(GPR35 receptor)in HT-29 cell as target,GPR35 receptor agonist zaprinast(1μmol/L)as positive control,DMR response value as the detection index,the agonistic and desensitizing activity of each fraction(100μg/mL)on GPR35 receptor was screened by label-free integrated pharmacological method,so as to screen active anti-allergic asthma fraction.HPLC-Q-TOF-MS method was used to identify the chemical composition of the selected active fractions.RESULTS:The alkaloids-free part of Ephedrae Herba was divided into two parts,involving the precipitated part before solid phase extraction and the 95%methanol elution part;from them,20 fractions were screened.Among them,the precipitated fraction F1.5-F1.10 and 95%methanol eluted fraction F2.5-F2.10 had a strong agonistic activity on GPR35 receptor;at the same time,GPR35 receptor agonist zaprinast showed a relatively strong desensitization activity.The signal intensity of DMR induced by F1.5-F1.10 in the precipitated part of HT-29 cells was even higher than that of reference drug zaprinast.By HPLC-Q-TOF-MS analysis,24 chemical components were identified from active fractions,involving 14 flavonoids,2 volatile oils,7 organic carboxylic acids,1 anthraquinones.CONCLUSIONS:The alkaloid-free part of Ephedrae Herba is mainly flavonoids and has anti-allergic asthma activity.

关 键 词：麻黄 非生物碱部位 G蛋白偶联受体35 过敏性哮喘 化学成分 鉴定 

分 类 号：R284.1[医药卫生—中药学]