华蟾素注射液中可疑致敏成分筛选的实验研究

目的:明确华蟾素注射液中可疑致敏原成分,为上市中药注射剂过敏原的筛查提供新的实验方法。方法:首先在临床上收集华蟾素注射液过敏患者血清;同时将华蟾素注射液中的可疑致敏原与牛血清白蛋白偶联合成人工抗原,在体外重现华蟾素注射液在人体内的过敏反应过程,利用血清学检测及激光共聚焦显微镜技术,检测RBL-2H3细胞被诱导脱颗粒释放组胺及TNF-α浓度的变化,与华蟾素注射液人体内过敏反应所产生的检测结果进行比对。结果:RBL-2H3肥大细胞组胺释放率、组胺及TNF-α浓度测定显示,脂蟾毒配基-BSA组、过敏患者血清组(用药后)组与标准对照组相比,差异均无显著性;华蟾酥毒基-BSA组、空白对照组与标准对照组相比,差异均有显著性(P<0.05)。结论:临床上收集的华蟾素注射液过敏患者血清中的过敏原成分应为脂蟾毒配基。

Experimental study of suspicious sensitization component screening on Huachansu injection

Objective: To explore a new experimental method for screening of allergens in post-market traditional Chinese medicine injections by confirming allergens in Huachansu injection.Method: First of all,the serum of patients allergic to Huachansu injection were collected,at the same time,the dubious allergen was conjugated to bovine serum albumin(BSA) by EDC coupling procedure to form complete antigen(BNP-BSA),which makes it possible to reproduce the allergic reaction of Huachansu injection in vitro.The histamine liberation ratio,the level of TNF-α and Histamine released from RBL-2H3 mast cell were detected;the above data were compared with those obtained in vivo.Result: The difference of the histamine liberation ratio,the levels of TNF-α and histamine of the resibufogenin-BSA group,group of patients allergic to Huachansu injection were not significant compared with those of normal control group.However,there were significant difference in those data among the cinobufagin-BSA group,the blank control and normal control group(P<0.05).Conclusion: The allergen in the serum collected from patients allergic to Huachansu injection is resibufogenin.