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
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篇名: | 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 |
TITLE: | Analysis of Chemical Components of Anti-allergic Asthma Active Fractions in Alkaloids-free Part of Ephedrae Herba |
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UnitaryC18柱和EclipseXDB-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/mL)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。 |
ABSTRACT: | 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 AC 18)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 C 18 column and Eclipse XDB-C 18 column. Using high expression G protein coupled-receptor 35(GPR35 receptor)in HT- 29 cell as target ,GPR35 receptor agonist zaprinast (1 μmol/L)as positive control ,DMR response value as the detection index ,the agonistic and desensitizing activity of each fraction(100 μg/mL)on GPR 35 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 F 1.5-F1.10 and 95% methanol eluted fraction F 2.5-F2.10 had a strong agonistic activity on GPR 35 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. |
期刊: | 2020年第31卷第09期 |
作者: | 靳京妹,于大永,侯滔,张秀莉,史丽颖 |
AUTHORS: | JIN Jingmei ,YU Dayong ,HOU Tao,ZHANG Xiuli,SHI Liying |
关键字: | 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 |
KEYWORDS: | Ephedrae Herba ;Alkaloids-free part ;G protein coupled-receptor 35;Allergic asthma ;Chemical component ; |
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