Ⅰ Type Hypersensitivity Study In Guinea Pigs Caused By Acanthopanax Antigen

Background:Chinese medicine injections in recent years, with clinical application used more widely, allergic reactions occurrence increased. As Chinese medicine injection drugs is a kind of emerging products, its composition is complex and not fully detected among the various components as well as the content amount. The allergic reactions have occurred in some cases. Among which type I allergic reactions is the most common and dangerous. In recent years, China has increasing emphasis on Chinese medicine injections which cause allergic reactions, and many have been reported. But the report mostly came out from PCA or ACA experiments in order to observe its evaluation of the main allergy symptoms. This method is subjective, which is difficult to quantify and standardize.ObjectivesThis study established for evaluation the type I allergic on the guinea pig. By using the herbal extracts from Acanthopanax antigenic substances. This study has proved the allergy reaction caused by the Acanthopanax and verify the presence of antigen. The positive results further validate the validity of the model design. Meanwhile medicine antigenic material sensitized with adjuvant required a preliminary comparison.Methods:1. Acanthopanax antigen preparation and protein detection:by simulating Acanthopanax Injection "boiled alcohol precipitation" of the product ion process to preparation Acanthopanax antigens. Add500g chopped herbs Acanthopanax in the water, boil it under high pressure, collect the active ingredients by repeat filtration, add ice ethanol precipitation to impurities, drying it by vacuum at room temperature to recovery ethanol, and obtain the resulting powder of Acanthopanax antigen6.582g. The content of protein amount is42μg/mg by assaying it with the Bradford method.2. The establishment of an allergic reaction model and grouping:The guinea pigs were divided into normal saline group(NS), OVA group, Acanthopanax Acanthopanax Freund’s adjuvant(F/Ag) and aluminum adjuvant group(AL/Ag) of4groups,8/group, each group were immunized four times after the first immunization booster once every two weeks.3. In vivo Stimulate:7days after the last booster immunization. injecting corresponding immunogen by hindlimb’s vein to stimulate. Observing each guinea pig’sreaction in the after3hours. Take the peritoneal lavage fluid(BLAF). Use ELLISA kit to detect the samples allergic mediators of histamine and leukotriene C4levels.4. Stimulate the in vitro cell excitation:7days after the last booster immunization, Take the peritoneal lavage fluid and peripheral blood. Separate the mass cell and mononuclear cell. in vitro stimulus and develop30mins by corresponding immunogen and then keep the supernatant. Use ELLISA kit to detect the samples allergic mediators of histamine and leukotriene C4levels.Results:1. After stimulate the in vitro,OVA group, Acanthopanax Acanthopanax Freund’s adjuvant and aluminum adjuvant group were all appeared conspicuous allergy symptoms. But there were no differences between Acanthopanax Acanthopanax Freund’s adjuvant and aluminum adjuvant group in allergy symptoms’s socres.2. After stimulate the In vitro, the histamine levels of BALF, the OVA group and Acanthopanax Freund’s adjuvant group were significantly different with control group(P<0.05). Although the histamine levels of AL/Ag group were raise, it has no significant difference with NS group;LTC4’s level, OVA group, Acanthopanax Acanthopanax Freund’s adjuvant and aluminum adjuvant group leukotriene levels were significantly higher compared with the control group (P<0.01).3. The supernatant of mononuclear cell of peripheral blood stimulate the in vitro, Acanthopanax Acanthopanax Freund’s adjuvant and aluminum adjuvant group histamine and leukotriene C4levels were all developed and were significantly higher compared with the control group(P<0.05; P<0.01).4. The supernatant of mass cell of peritoneal lavage fluid stimulate the in vitro, Acanthopanax Acanthopanax Freund’s adjuvant and aluminum adjuvant group histamine and leukotriene C41evels were all developed and were significantly higher compared with the control group(P<0.05; P<0.01).Conclusion:1. Acanthopanax’s extract of water boiling and precipitation with ethanol can cause type Ⅰ allergic reaction in guinea pigs. It have allergenicity. the production of traditional Chinese medicine injection process need to be improved. The current purification methods can not be totally remove the antigenic material of Chinese herbal medicines;2. Type I allergic guinea pig model can evaluate traditional Chinese medicine injections. Detecting the allergic mediators to evaluate aensitization safety of Chinese medicine can quantification.3. Freund’s adjuvant and aluminum adjuvant have no differencese in causing type I allergic reaction in guinea pigs. Aluminum adjuvant are also available in the evaluation of traditional Chinese medicine injections.