Optimization Of Anaphylactoid Reaction Models And Application In The Detection Of The Traditional Chinese Medicine Injections By Anaphylactoid Models

In recent years, adverse reactions of the traditional Chinese medicine injection (TCMI) increased dramatically and the safety of it is becoming a major concern of Chinese medicine. Immediate hypersensitivity is one of the most complex and the key problems of TCMI. Immediate hypersensitivity includes type I allergic reaction and anaphylactoid reaction. In the clinical allergic reactions, the incidence of anaphylactoid reactions accounts for about77%of the total number. How to establish the accurate and fast detecting system is one of most important issue of allergic sensitization research.Anaphylactoid reaction is a common kind of acute allergic reactions. Anaphylactoid reaction is through activating mast cells/basophils directly or activating the complement system, it makes the target cells degranulated and release allergy mediators, therefore, it can cause local or systemic allergy symptoms. In this study, the models of rats and dogs were used to study complement activation related anaphylactoid. And the skin vascular permeability in mice, which mechanism is direct stimulation of mast cells/basophils, was used to test in the direct pathways. These anaphylactoid models were all used to test TCMIs, some monomers and excipients.1. Optimization and application of anaphylactoid reaction model in ratObjective:In order to optimize the anaphylactoid model in rat, we selected two kinds of rats (Wistar and BN), and increased some characteristic indexes. Methods:The MAP was used to evaluate the sensitive in BN and Wistar rats. The complement factors Bb and C4d were used to differentiate the pathways of complement activation. At the same time, this animal model was used to screening10kinds of TCMIs,5 kinds of monomers and3kinds of excipients. Results:The MAP of BN rats decreased more sharply than that of Wistar rats. Shuanghuanglian Injection can make the MAP decrease significantly, the content of histamine increase and IgE unchanged. Conclusions:BN rats were more sensitive than Wistar rats. Shuanghuanglian Injection has obvious anaphylactoid reactions. Tween80has the characteristics of complement activation, and it may activate the classic and alternative pathways of the complement system. The anaphylactoid model of BN rats is suitable of testing for TCMIs.2. Optimization and application of anaphylactoid reaction model in Beagle dogObjective:We increased some indexes of anaphylactoid model in dogs, analysed the characteristic indexes, in order to discuss the mechanism of anaphylactoid. Methods: Except for usual hematology indexes, such as histamine, IgE, SC5b-9, we increased the complement parts of Bb and C4d and increased detection of the complement pathways. Based on the physiological indexes (MAP and HR), ECG and PAP were added to analyse the cardiovascular changes. And these anaphylactoid indexes were all optimized in the model. At the same time, the animal model was used to test10kinds of TCMIs,5kinds of monomers, and3kinds of excipients. Results: Shuangguanglian Injection, Tween80and CrEL can make the MAP decrease, the content of histamine increase and IgE unchanged. Conclusions:Shuangguanglian Injection, Tween80and CrEL had obvious characteristics of anaphylactoid reactions. CrEL and Tween80had the obvious characteristics of complement activation, and they both may activate the classic and alternative pathways of the complement system.3. Optimization and application of anaphylactoid reaction model in miceObjective:Building a more stable and reliable mouse model through selecting the optimal experimental conditions. Methods:The content of Evans blue was screeninged in order to optimize the model. H1blockers diphenhydramine and mast cell membrane stabilizer cromolyn sodium were used to further study the mechanism of skin vascular permeability. Results:The diphenhydramine and cromolyn sodium can decrease the production of blue stain. And there are positive results caused by Shuanghuanglian Injection and Yinzhihuang Injection. Conclusions:Vascular permeability in mice can make the experimental results reliable and reproducible under optimal conditions.4. Establishment of hemolysis test and complement activation test in vitroObjective:The stimulation of complement in blank serum was researched in order to further study the pathway of complement activation. Methods:The drugs were maxed with the blank serum of dogs and humans, the hematological index in the sera were detected by ELISA. Through the complement blocking agents, EGTA/Mg2+, sCRl, the mechanism of complement activation was researched. Results:Tween80can activate the complement system, and activate the classic and alternative pathways. Conclusions:This model is suitable to analyse the complement pathways of the anaphylactoid reactions.