中国中药杂志

2019, v.44(08) 1588-1595

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基于免疫指纹图谱法筛查双黄连注射剂的致敏成分
Screening of Shuanghuanglian Injection allergenic ingredients based on immune fingerprint

文雯;贺福元;刘文龙;周逸群;刘红宇;
WEN Wen;HE Fu-yuan;LIU Wen-long;ZHOU Yi-qun;LIU Hong-yu;Department of Pharmaceutics,Hunan University of Chinese Medicine;First Affiliated Hospital of Hunan University of Chinese Medicine;Hunan Provincial Key Laboratory of Druggability and Preparation Modification of Traditional Chinese Medrcine;

摘要(Abstract):

该文采用免疫指纹图谱法,以酶联免疫吸附法(ELISA)结合HPLC/MS法筛查双黄连注射剂(SHLI)的致敏性成分,制备SHLI的鼠抗兔血清免疫球蛋白E抗体(抗Ig E抗体),通过ELISA包埋抗Ig E抗体可成功吸附SHLI及其不同给药途径的含药血浆样品中的致敏原,提示SHLI可引起大鼠产生Ⅰ型超敏反应。建立抗Ig E抗体吸附前后各样品的HPLC指纹图谱和质谱图,由HPLC指纹图谱的相似度和MS图谱归纳结果可知,不同给药方式可改变SHLI的致敏性,SHLI及其含药血浆中可被特异性抗Ig E抗体吸附的成分有22种,多为酸类、酯类及含氮化合物,基于超分子理论推断这些酸类、酯类及含氮化合物来源于SHLI或机体,可能形成超分子半抗原,致使其口服不过敏而作为注射剂使用时具有免疫毒性引发过敏反应。比较传统的中药注射剂致敏原筛查方法,免疫指纹图谱法不仅限于单成分致敏原筛查,该方法纳入成分更全面,灵敏度高,操作方便,可为今后中药注射剂过敏反应的研究方法提供参考。
In this paper,immune fingerprint was used to screen the allergenic components of Shuanghuanglian Injection(SHLI) by enzyme-linked immuno sorbent assay(ELISA) combined with HPLC/MS method. ELISA-embedded anti-IgE antibody could successfully adsorb allergens in SHLI and its plasma samples containing drugs through different routes of administration,suggesting that SHLI can induce type I hypersensitivity in rats. HPLC fingerprints and MS map of SHLI and drug-containing plasma samples from different routes of administration before and after anti-IgE antibody adsorption were established. According to the similarity evaluation of HPLC fingerprints and analysis results MS map,the sensitization of traditional Chinese medicine injections can be changed by different administration methods. There were 22 kinds of components that can be adsorbed by specific anti-Ig E antibodies in Shuanghuanglian Injection and its drug-containing plasma,most of them were acids and nitrogen compounds. Based on supramolecular theory,it was inferred that these compounds came from SHLI or body,and may form supramolecular hapten,which results in immunotoxicity and allergic reaction when being used as injection instead of oral liquid. Immune fingerprint is not only used to screen out single component allergen,but also more comprehensive,sensitive and easy to operate. It can provide reference for the future research methods of allergic reaction of traditional Chinese medicine injections.

关键词(KeyWords): 双黄连注射剂;免疫指纹图谱;超分子;过敏反应;抗IgE抗体
Shuanghuanglian Injection;immune fingerprint;supramolecular;anaphylactoid;anti-IgE antibody

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年项目(81803729);; 湖南省自然科学基金项目(2016JJ2097)

作者(Author): 文雯;贺福元;刘文龙;周逸群;刘红宇;
WEN Wen;HE Fu-yuan;LIU Wen-long;ZHOU Yi-qun;LIU Hong-yu;Department of Pharmaceutics,Hunan University of Chinese Medicine;First Affiliated Hospital of Hunan University of Chinese Medicine;Hunan Provincial Key Laboratory of Druggability and Preparation Modification of Traditional Chinese Medrcine;

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DOI: 10.19540/j.cnki.cjcmm.20190325.301

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