Experimental Study On Anaphylactoid Reactions Of Different Dosage Forms Of Traditional Chinese Medicine

1 Purpose and significanceFirstly, the purpose of this study is to select the Chinese HowNet database for analysis the traditional Chinese medicine and literature on allergic diseases, then collate and summarize the 11 kinds of traditional Chinese medicine injections on the sex, age, type of allergic reaction, reports and other information.Secondly, through the questionnaire survey, analysis residents cognition of allergy and anaphylactoid reaction, and then screened by drugs of mast cell and RBL-2H3 cell model, determine the two model which is more sensitive, selected the forms of Chinese medicine which cause obvious anaphylactoid effect, and then carry out the body allergic sensitization, preliminary quasi sensitive analysis of three kinds of dosage form in traditional Chinese medicine anaphylactoid reaction.The significance of this study lies in two kinds of anaphylactoid reaction by in vitro cell model experiment, to determine which cell model is sensitive, then screening the anaphylactoid experiment on different formulations of traditional Chinese medicine, evaluate the allergic type caused by dosage form of commonly used traditional Chinese medicine, combined with the result of the literature and questionnaire, then conduct the sensitivity evaluation to provide the reference for the clinical the results,which has practical application value in improving the prediction of the traditional Chinese medicine clinical anaphylactoid reaction. 2 Methods 2.1 Literature and questionnaire of traditional Chinese medicine allergic diseasesFirst of all, through the literature analysis the four types of allergic hypersensitivity disease, then using retrospective study method to collate and summarize the 11 kinds of traditional Chinese medicine injections on the sex, age, type of allergic reaction, reports and other information. Then, based on the range of people whose age are above 15, to investigate whether people who widely used the traditional Chinese medicine have enough understanding of its adverse reactions, and based on the data of questionnaire to analyze the cognition of people on allergic and anaphylactoid reaction. 2.2 Selection of cell lines and study on their characterizationsThe research studied on the growth characteristics on mast cells and RBL-2H3 cells, which provide the prerequisite for the different dosage of TCM anaphylactoid reaction experiment. The growth curves of two cells with different concentrations were drawing by continuous cultivation method and the MTT assay, which we could find the best experimental cell number and time for detection. 2.3 Different forms of traditional Chinese medicine anaphylactoid reaction screeningUse the method of MTT to detect the influence of different concentrations of injections of traditional Chinese medicine, Chinese medicine granules and decoction of Chinese medicine respectively used on the mast cells and RBL-2H3 cells. Screening out the sensitive cell line of three forms of traditional Chinese medicine in common, then selected the forms of severe anaphylactoid reactions caused by traditional Chinese medicine through comparative analysis, preparing for the next step of the experiment in vivo. 2.4 Different forms of traditional Chinese medicine in vivo anaphylactoid sensitizationThrough allergic anaphylactoid experimental screening of different forms of Chinese medicines on mast cells and RBL-2H3 cells, selected out that Shuanghuanglian injection, Qingkailing granules and Houttuynia liquid caused the strong effect. To investigate the characteristics of in vivo function, we inject Shuanghuanglian on mouse tail vein, Qingkailing granules and Houttuynia liquid by intragastric administration to explore whether they can stimulus the anaphylactoid effects, and then through the detection of skin locus coeruleus area and observe the mouse behavior changes of different dosage forms of Chinese traditional medicine, preliminary analysis the anaphylactoid sensitization in vivo function. 3 Results 3.1 The result of literature and questionnaire of traditional Chinese medicine allergic diseasesThrough the statistics and case retrieval of drug allergy, finding that drug allergy mainly cause by injection, and 11 kinds of traditional Chinese medicine injections in 1352 cases, many of which type I allergic reaction, a total of 1281 cases, accounting for 94.75%; part of traditional Chinese medicine injections can also cause II, III, IV type hypersensitivity reactions, and in general, allergic reaction in males accounted for 54.22%.Through the questionnaire survey for the residents of the cognition of allergy first and allergy again, found that the vague knowledge of resident’s cognition of allergy and anaphylactoid was cognitive, clearer and the proportion of 18.19%, the vast majority of people are aware of the less. And the residents for the first time in the recognition of anaphylactoid, first clear the lowest proportion are 10.77%. 3.2 Selection of cell lines and study on their characterizationBy drawing the cell growth curve, we found that mast cells have the best proliferation activity with the inoculation concentration of 1.0×105 cell/m L; RBL-2H3 cells have the best proliferation activity with the inoculation concentration of 1.0×105. Mast cells lines proliferated in 0h~24h, and had the best cell viability at 24 h, so we could determine24 h as the time node for mast cells drug experiments.RBL-2H3 cells lines proliferated rapidly in 24h~72h, and had the best cell viability at 72 h, then reached a plateau, so we could determine72 h as the time node for RBL-2H3 cells drug experiments. 3.3 Different forms of traditional Chinese medicine in vitro anaphylactoid reaction screeningThe anaphylactoid reaction screening results show that Shuanghuanglian injection,Qingkailing granules and Houttuynia, liquid can cause strong anaphylactoid reaction(p<0.05). Comparison of three different dosage forms of traditional Chinese medicine on mast cells and RBL-2H3 cells of the anaphylactoid effect, the results show that the anaphylactoid effect caused by Shuanghuanglian injection is the best which IC50 is 35.553 L/m L for 1h, and the sensitive cell lines effects caused the corresponding to anaphylactoid reaction is the mast cells. 3.4 Three forms of traditional Chinese medicine in vivo anaphylactoid sensitizationThrough comparing the decoction results of Shuanghuanglian injection, Qingkailing granules and Herba Houttuyniae anaphylactoid hypersensitivity in vivo experiment, observed the behavioral changes and detection of mouse skin blue spot area, found that three forms of traditional Chinese medicine anaphylactoid reaction was not obvious, but administered again after sensitization, Shuanghuanglian injection group showed obvious, Qingkailing and Houttuynia group had no obvious change. 4 Conclusions 4.1 First, the search of literature of Chinese medicine allergic diseases were shocking, which showed that the field of medicine, hospital and community universal should payed more attention to, in addition to some traditional hypersensitivity, also special emphasis on anaphylactoid reactions caused by the first drug case; Second, through the questionnaire survey found that the cognition of anaphylactoid of residents is low, should strengthen the propaganda of Chinese medicine knowledge, and improve the cognition of residents of Chinese medicine, in order to understand the Chinese medicine correctly. 4.2 Through the two cell lines in vitro growth law, we grasp the best inoculation of cell concentration and cell growth duration, which provides the premise to study different dosage forms of traditional Chinese medicine in vitro anaphylactoid reaction. 4.3 Through different forms of traditional Chinese medicine in vitro anaphylactoid reaction experiment, we found that two kinds of cell lines to different dosage forms of Chinese traditional medicine with different sensitivity. The strongest corresponding caused by Shuanghuanglian injection when it’s IC50 is 35.553 L/m L on mast cells. 4.4 By inducing the in vivo experimental sensitivity with three different dosage forms of Chinese traditional medicine, comparied the experimental results of Shuanghuanglian injection, Qingkailing granules and Herba Houttuyniae anaphylactoid hypersensitivity in vivo. Observed the changes in the behavior of mice, anaphylactoid process performance is not obvious, but administered again after sensitization, Shuanghuanglian injection showed obvious, mice without abnormal behavior of Qingkailing and Houttuynia cordata.