中药注射剂致敏成分筛查方法的研究

Study on screening method of allergenic ingredients of TCM injections

目的:本文以血栓通注射液为例,筛选其内在致敏原,并试图探寻一种系统、可靠的中药注射剂致敏成分筛查方法,为其它中药注射剂致敏成分筛查提供借鉴和参考。方法:利用P815细胞体外脱颗粒模型对血栓通注射液分离成分进行过敏筛查;通过小鼠直接腘窝淋巴结实验模型对血栓通注射液分离成分进行过敏筛查验证;分析并结合此两种筛查方法试验结果,综合评判血栓通注射液内在成分的致敏潜能。结果:P815细胞脱颗粒模型:血栓通注射液、三七总皂苷、人参皂苷Rb1组胺释放量、β-氨基己糖酶释放率和细胞脱颗粒率三项指标均有显著性差异(P<0.01),人参皂苷Rg1组胺释放量、脱颗粒率有显著性差异(P<0.05);直接腘窝淋巴结实验模型:三七总皂苷组、单体人参皂苷Rb1及人参皂苷Rg1均WI>2,CI>5。结论:人参皂苷Rb1和Rg1可能是导致血栓通注射液临床过敏的内在因素。建议:结合体内外两种过敏筛查模型,综合评判中药注射液致敏潜能,探寻一种由体外细胞实验初步筛选到体内动物实验进一步验证的更加可靠、有效的中药注射剂致敏成分筛查方法。

Objective： In this paper we take Xueshuantong injection for example, screening its inherent allergens, and trying to establish a systematic and reliable allergenic ingredients screening methods of TCM injections. In order to provide references for screening allergenic ingredients of other TCM injections. Methods： Using P815 cell degranulation model to screen components of Xueshuantong injection. By means of direct popliteal lymph node assay to verify allergenic ingredients of Xueshuantong injection components. Analyzing and combining these two test resuhs, to evaluate sensitizing potential of inherent components of Xueshuantong injection comprehensively. Results： P815 cell degranulation model： Histamine release, β- amino hexose enzyme release rate and cell degranulation rate, this three indicators of Xueshuantong injection group , PNS group and ginsenoside Rbl group have significantly difference （ P 〈 0.01 ）. Histamine release and degranulation rate of group ginsenoside Rgl has significant difference （ P 〈 0.05）. Direct popliteal lymph node assay model： Groups of PNS, ginsenoside Rbl and ginsenoside Rgl all have WI 〉 2, CI 〉 5. Conclusion： The internal factors of clinical allergy of Xueshuantong injection may relation to Ginsenoside Rbl and Rgl. Recommendation： Combining this two models, to evaluate sensitizing potential of TMC injection comprehensively. Exploring a systematic and reliable allergenic ingredients screening methods in which including the in vitro cell experiments to preliminary screening and in vivo animal experiments to further validate the allergenic ingredients of TMC injections.