The Development Of BALB/c Mice Anaphylaxis Model And Its Application In The Detection Of The Anaphylaxis Induced By Traditional Chinese Medicine Injection

Traditional Chinese Medicine Injection (TCMI), a new preparation, is originally created in our country. It has the characteristics of convenient administration, high bioavailability and fast-acting. With the extensive application of TCMI, the adverse drug reactions happen frequently, especially allergic reponses, arousing the public attention. Because the complicated composition of TCMI, its potential allergens may be associated with macromolecules as well as small molecules, or both of them may be related, which claim higher demands for the development of anaphylaxis animal models.The species most commenly favoured with respect to anaphylaxis animal models is the BALB/c mice. This is largely driven by the availability of their small sizes, short breeding cycle as well as various immunological and molecular reagents. Moreover, many aspects of immune regulation similar mechanisms are shared between men and mice. In addition, BALB/c mice are predisposed to be high IgE responders, and tend to Th2reactions. Therefore, they can better simulate the clinical occurrence of allergic reactions.In this study, we took BALB/c mice as experimental subjects; tried to create a type of anaphylaxis animal model which could be used for the detection of macromolecules, but also for small molecules, which could dismiss the species differences effectually and improve the accuracy and reliability of detection. And then, we made use of the above model developed to detect the sensitization potential of QKLI and GGSI in order to find certain suitable animal model, providing experimental basis for the pre-clinical evaluation of anaphylaxis induced by TCMI. In the first part of this study, we tried to improve the sensitized procedure by optimizing the route of antigen exposure, frequency, dose and antibody detected time. The indicators of anaphylaxis including total IgE levels, total IgG levels, specific IgE levels, spleen cell cytokine levels, systemic symptoms and so on are also determined in order to develop the BALB/c mice model for the detection of macromolecules. After the development of the above model, three proteins with different sensitizabilities were applicated in the evaluation of the BALB/c model developed. Finally, we applied this BALB/c mice model for macromolecules in the detection of anphylaxis induced by QKLI, GGSI and related protein extracts of QKLI. The optimal sensitized methods chose were:BALB/c mice were sensitized subcutaneously with0.5mg/mouse of OVA. Seven days later the treatment was repeated. Mice were exsanguinated or challenged14days after the initiation of exposure. The suitable indicators of anaphylaxis included specific IgE level, spleen cell cytokine levels, and systemic symptoms. The study found that the allergic reactions of buffalo horn protein extract and gardenia protein extract from QKLI were positive.In the second part of this study, we took D-penicillamine hydrochloride (D-pen) and streptozotocin (STZ) as positive control drugs, phenobarbital (PB) as negative control drug. Dose-respose relationship and kinetics of PLN response of positive control drugs were studied to construct the BALB/c mice direct popliteal lymph node assay (d-PLNA) and secondary PLNA (s-PLNA) models. Finally, we applied this BALB/c mice PLNA model in the detection of anphylaxis induced by QKLI, GGSI as well as their major monomer components. In accordance with the positive standard, mass index (MI)≥2and cell index (CI)≥5, the lowest effective dose of D-pen and STZ were both0.75mg/mouse, the optimum examing time was the seventh day after D-pen and STZ were given. And then BALB/c mice d-PLNA and s-PLNA models were constructed under the conditions obtained above. The study found that there may be some small molecular allergic ingredients in the QKLI, and allergic reactions could be induced when these ingredients reached a certain concentration.In a word, this BALB/c mice model was able to roundly, accurately detect the sensitized potential of QKLI and GGSI, which was expected to be further applied in the detection of anaphylaxis induced by other TCMIs.