IgG-HepG2细胞评价复方苦参注射液及其生产过程可能产生致敏原的研究——利用生物技术进行过程监控致敏性的方法学探索  

Evaluation of allergen during processes of Fufang Kusheninjection by IgG-promoter-HepG2 cell assay in vitro

作　　者：袁梽漪[1] 王玉刚[1] 雷帆[1] 卢希[1] 赵爽[1] 姜敬非[1] 李慧玉[1] 陈勃杭[1] 王智民[2] 邢东明[1] 丁怡[1] 杜力军[1] 

机构地区：[1]清华大学医学院生命科学学院教育部蛋白质重点实验室,北京100084 [2]中国中医科学院中药研究所,北京100700

出　　处：《中国中药杂志》2011年第14期1850-1854,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30801523;30973896;81073092);国家"重大新药创制"科技重大专项(2009ZX09308-003;2009ZX09502-021;2009ZX09502)

摘　　要：目的:利用IgG promoter-HepG2细胞对复方苦参注射液生产过程中可能产生致敏原的生产节点进行研究。方法:构建IgG启动子调控绿色荧光蛋白表达质粒,转染入HepG2细胞,与常见致敏性物质(葛根素,卵白蛋白,LPS和伤寒菌疫苗)、复方苦参注射剂中使用到的辅料(NaOH,醋酸,聚氧乙烯脱水山梨醇单油酸酯即吐温80和乙醇)和生产过程中关键节点样品(S1～S8)进行孵育,30 m in后拍照,图像分析软件统计细胞荧光强度变化。结果:IgG promoter-HepG2细胞对常见致敏性物质呈阳性反应。注射剂辅料中醋酸和吐温80有致敏性,生产过程8个样品中,2个样品有显著致敏性。结论:IgG promoter-HepG2细胞对常见致敏原反应敏感,特异性高,适用于中药注射剂中致敏原快速筛选,可实现对中药注射剂生产过程中致敏原的实时监控。Objective： To study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro. Method： By transfecting a IgG promoter-regulating the expression of green fluorescent protein（GFP） plasmid into HepG2 cells, this transferred cells were incubated with common allergens （like puerarin, ovalbumin, LPS or Sal typhoid vi polysaccharide vaccine）, excipients using in Fufang Kushen injection（ NaOH, acetic acid, Tween-80 and ethanol） and samples from the key production processes of the injection for 30 minutes. Fluorescent photographs were analyzed the fluorescence intensity of the cells by using an image analysis software. Result： All of common allergens significantly increased the IgG expression. Two of the excipicents, acetic acids and Tween-80 were shown to increased the IgG expression, while others had no effect on IgG expression. In the 8 samples from the key processes in the production of Fufang Kushen injection, two of them stimulated IgG expression. Conclusion： IgG promoter-HepG2 cells are highly sensitive and specific to allergens, and thus can be applied to rapid screening of allergens in components and injections in transcriptional level. It is possible to use the IgG-promoter HepG2 cells in a real-time monitoring of allergens in the production processes of Chinese medicine injections.

关 键 词：IGG 中药注射剂 苦参 过敏反应 

分 类 号：R286[医药卫生—中药学]