细胞效应谱技术研究绿原酸类成分致类过敏反应的真实性

Would chlorogenic acids (CAs) cause the anaphylactoid reaction (AR)? The topic continues to be controversial. And it had already impeded the sound development of traditional Chinese medicine injections (TCMJs) containing CAs. Those problems mainly focus on: (1) it is not clear about the substance contents and features of CAs and how CAs caused the AR when those acids existed in the whole context of TCMJs, such a complex system; (2) there is little sensitive and clinic-related method to detect the AR. Therefore, the project begins from the analysis of different substances of CAs, paying close attention to sensitive cells which cause the AR. Meanwhile, CAs and their catabolites, isomers, G-protein-coupled combinations-such CAs are targeted in the study. Basing on chemical features of substance contents, we introduce cyto-dynamics and living cell monitoring technology that is real-time, non-invasive and free label, to represent biological characteristics of interactions between CAs and AR-related cells (including cells from patients, healthy people, normal animals and standard cell bank). Then the established new method to detect the AR could be used to confirm the validity of the CAs-induced AR. We hope this method can provide a new technical support to improve the quality control level of TCMJs containing CAs and reduce clinical adverse reactions.

绿原酸类成分是否会引发类过敏反应？至今尚无确切定论，但已给含绿原酸类成分的中药注射剂持续健康发展蒙上了一层阴影。问题的聚焦点在于：（1）绿原酸类成分物质内涵及其在注射剂中的本底信息不明确；（2）缺乏灵敏且关联临床的类过敏反应检查方法。为此，本项目从绿原酸类成分不同物质内涵的解析入手，关注引发类过敏反应的敏感细胞，以绿原酸及其分解产物、异构体、蛋白偶联结合体等绿原酸类成分为研究对象，在物质内涵化学表征的基础上，引入细胞动力学，采用非侵入、无标记、实时在线的活细胞监测技术，表征绿原酸类成分与类过敏反应相关细胞（含临床患者源、健康人源、正常动物源、标准细胞库源细胞）相互作用的生物学特性，建立类过敏反应检查新方法，辨识绿原酸类成分致类过敏反应的客观真实性，以期为提升含绿原酸类成分中药注射剂的质量评控水平，减少临床不良反应发生提供新的技术支持。

绿原酸是否为类过敏原备受争议，分析其可能是缘由，与绿原酸本身的稳定差及在体内易转化有关，同时也缺乏灵敏、实时表征技术。为此，研究中建立了具有非侵入、无标记、实时在线等技术优势的活细胞监测技术，可用于表征在接近生理或病理条件下监测可疑成分致肥大细胞脱颗粒效应程度。研究发现，细胞指纹谱技术可较好的表征不同绿原酸类成分（10种）致肥大细胞脱颗粒效应；绿原酸在一定条件下转化为脱颗粒作用更强的隐绿原酸和新绿原酸引发类过敏反应，而隐绿原酸和新绿原酸难以转化为绿原酸；进一步采用蛋白空间结构分子虚拟对接技术，也印证了这一结果。这一发现对回答绿原酸是否为类过敏原以及提升含绿原酸的注射剂质量有着重要的参考价值。. 相关研究成果在Anal Bioanal Chem、Sci Rep、J. Sep. Sci、《药学学报》发表论文4篇；中国专利1项；培养研究生3名，参加学术交流会议2次。