The Development Of Anaphylaxis Animal Model And Its Application In The Detection Of The Anaphylaxis Induced By Shuanghuanglian Injection

The anaphylaxis animal model is essential in the study of anaphylacxis induced by medicines, because the animal model is the prerequisite for screening for allergens and the basis for preventing allergic diseases in clinic. However, the current anaphylaxis animal models are generally lack of sensitivity and accuracy, which causes negative results in predicting clinical allergic diseases and poses difficulties on the anaphylaxis study. As a noval form of Traditional Chinese Medicine Injection (TCMI), Shuanghuanglian Injection (SHLI) recently has induced many serious anaphylaxis reactions, arousing the public attention. There are many factors to induce the frequent occurance of SHLI-induced anaphylaxis reactions, such as the complex composition, the failure of quality control, the inaccuration of allergy tests, which claims a high pay for the development of anaphylaxis animal model.In this study, we optimized the experimental protocols and screen for detection indicators to develop rats, guinea pigs and mouse anaphylaxis animal models. Then we made use of these animal models in the detection of anaphylaxis induced by SHLI in order to find some available animal model, providing experimental basis for the pre-clinical evaluation of anaphylaxis induced by TCMI.1. The development of rats model and the application in the detection of SHLIAim:To find the allergy-sensitive rat species, develop a comprehensive detection system and apply in the detection of anaphylaxis induced by SHLI. Methods: Compare the allergy-sensitivity of Wistar. SD and BN rats in the detection of anaphylaxis. After the combination of aluminum hydroxide adjuvant, the route of subcutaneous injection and the five times senstitization, we tried to improve the detection sensitivity. We also determined the feasibility of total IgE levels, specific IgE levels, IL-4, the local and systemic symptoms, the allergy mediators, the blood pressure and the pathology examination as indicators of anaphylaxis. Finally, we applied this rats model in the detection of anaphylaxis induced by SHLI. Results:BN rats were more sensitive in anaphylaxis detection than Wistar rats and SD rats, being able to detect less OVA. When improved, the BN rats model could detect as less as 0.1 mg OVA. This BN rats model could detect the anaphylaxis induced by SHLI in total IgE, specific IgE, IL-4, symptoms and mediators. Conclusion:With these indicators, this BN rats model was able to accurately, sensitively and comprehensively detect tiny allergens as well as SHLI, which is expected to be further applied to the anaphylaxis detection of other TCMIs.2. The develoment of guinea pigs model and the application in the detection of SHLIAim:To develop a sensitive and quantitative guinea pigs model and apply in the detection of anaphylaxis induced by SHLI. Methods:On the basis of the guinea pig active systemic anaphylaxis (ASA) assay, we added the allergy mediator detection and pathology examination as indicators. We also determined the optimized challenge time by antibody level detection in order to elevate the detection senstitivity and objectivity. Finally, we applied this guinea pigs model in the detection of anaphylaxis induced by SHLI. Results:Compared with the mimimum detection line of 0.3 mg OVA. this guinea pigs model was able to detect tiny OVA, up to 0.01 mg. However, this guinea pigs model failed to demonstrate any severe allergic symptoms, moreover, this model failed to detect the increase of allergy mediator and the lung pathology change. Conclusion:The guinea pigs model is very sensitive to large molecule allergens, such as OVA, however this model failed to detect the anaphylaxis induced by SHLI, indicating the allergens in SHLI might be low molecule. This guinea pigs model may be used as supplementary of BN rats model in the detection of anaphylaxis induced by medicine.3. The development of mouse model and the application in the detection so SHLIAim:To develop a mouse model and apply in the detection of anaphylaxis induced by SHLI. Methods:We sensitized the BALB/c mouse via subcutaneous injection and then detected the antibody, observed the systemic reactions, conducted foot leakage experiment and examined the pathology change to judge the occurance and degree of anaphylaxis induced. Finally, we applied this mouse model in the detection of anaphylaxis induced by SHLI. Results:Only the BALB/c mouse in 1.0 mg OVA group showed positive results on the Day 21 but the 0.1 mg and 0.01 mg OVA group both showed negative results. Moreover, we got negative results in the detection of SHLI using this BALB/c mouse model. Conclusion:This BALB/c mouse model should be improved to increase the detection sensitivity, and currently this mouse model is not suitable as animal models to detect the anaphylaxis induced by SHLI.