基于活细胞指纹谱监测的中药注射剂过敏反应预警新方法研究

Although the clinical efficacy of Chinese herbal injection is well accepted, allergic reactions occur frequently. It is difficult for current quality standards to prewarn this kind of allergy. There are some limitations in the present allergic reaction detecting methods: (1) There are many disadvantages in animal experiments because of sensitive indicators lacking and poor correlation; (2) Chemical fixation and colour reaction are required in detection of allergy mediators or scanning/transmission electron microscopy , which are difficult to operate and might cause biological information loss; and lack of uncontinuous dynamic monitoring way would result in that only certain phase can be observed, instead of all phases; at the same time, Chinese herbal injection are often not apparent but colored, so it is easy to interfere determination results. Generally, allergic reactions are induced by immune cells. In this project, from the perspective of cytokintics, we will apply online monitoring technology with non-invasive, label-free and real-time characteristics to monitor living cells to characterize, using bio-response profile, the bioinformatic interactions between Shuanghuanglian injection (including qualified samples and clinical adverse reactions samples) and cells related to allergic reactions (including the cells of adverse reactions patients). Combining with real-time image monitoring, new prewarning method for allergic reaction of Chinese herbal injection will be established in this project, which is expected to provide a new technological support for the quality assessment control of Chinese herbal injection and the reduction of clinical adverse reactions.

中药注射剂临床疗效确切，但过敏反应频发，现行质量标准难以对其实现早期预警。过敏反应常规检测方法存在一定局限：（1）动物实验干扰因素多，缺乏灵敏指标，相关性不强；（2）过敏介质或扫描/透射电镜检测需化学固定和显色处理，操作繁琐且生物信息易丢失；只能观察某一时相状态，缺乏连续动态监测；同时中药注射剂一般具有色泽，易干扰检测结果。过敏反应多由免疫细胞介导引发，本项目拟从细胞动力学视角，采用非侵入、无标记、实时在线的动态活细胞监测技术，以活细胞指纹谱表征模式药双黄连注射液（含正常样品和临床不良反应样品）和过敏反应相关细胞（含不良反应患者源细胞）间相互作用的生物信息，辅以实时图像监测，建立中药注射剂过敏反应预警新方法，以期为提升中药注射剂质量评控水平、减少临床不良反应的发生提供新的技术支持。

本项目旨在建立中药注射剂过敏反应早期预警关键技术，提出生物活性指纹谱（bio-response fingerprinting）学术概念；从细胞动力学视角，采用非侵入、无标记、实时在线的动态活细胞监测技术，实现让细胞在接近生理或病理条件下，以活体状态进行细胞生物学特性监测模式药双黄连注射液（含正常样品和临床不良反应样品）和过敏反应相关细胞（含不良反应患者源细胞）间相互作用。结果发现，动态活细胞指纹谱更有利于监测反映产品质量信息，是现有以化学指纹谱技术为核心的评控手段的有益补充；还发现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；进一步研究发现绿原酸类成分可与靶细胞肥大细胞表面G蛋白偶联受体结合，引发肥大细胞脱颗粒。这一研究为下一步从分子机制角度阐释双黄连注射液中绿原酸类成分致类过敏的客观真实性及其作用机制提供了重要线索。.相关研究成果在Anal Bioanal Chem、Sci Rep、PLoS ONE、Chin Sci Bull、《药学学报》、《中国中药杂志》等发表论文18篇，主编著作2部；获国际专利授权2项、中国专利授权3项；成果载入国家标准（1项）、大学教科书（2部）、药学工具书（5部），并实现产业化转让2项；获省部级科研成果奖二等奖1项；共同组织中国科协第266次青年科学家论坛，参加国际学术交流会议2次，受到了同行专家的广泛关注；后续入选北京市卫生系统高层次卫生技人才、扬帆计划等；兼任国家级学会常务委员、省级学会主任委员、常务委员等。新近获批临床合理用药生物特征谱学评价北京市重点实验室。