An experimental study of combination of allergic and anaphylactoid reactions to Xuesaitong Injection(freeze-drying)based on RBL-2H3 cells and ICR mice model
Objective To establish an allergic and anaphylactoid superimposed reaction RBL-2H3 cells and ICR mice model for the study of allergic and anaphylactoid reaction to Xuesaitong Injection(freeze-drying)(XST).Methods This study employed the β-hexosidase and histamine release rate of RBL-2H3 cells as evaluation indices in vitro to determine the optimal dose of DNP-IgE,DNP-BSA,and the duration of interaction with C48/80.The study investigated the effects of various doses of XST on cell viability and degranulation after stacking.In vivo,ICR mice were used as experimental subjects,and the scores of(type)allergic reaction symptoms and the contents of immunoglobulin E(IgE),histamine,5-hydroxytryptamine,vascular endothelial growth factor A(VEGF-A),and terminal complement complex(SC5b-9)in plasma were used as evaluation indicators to screen positive conditions for OVA(2.5,5.0,10.0 mg·kg-1)and C48/80(1,2,4 mg·kg-1)allergy-like allergic models,and finally,the sensitization of XST(60,120,240 mg·kg-1)and whether it would cause allergic or allergic-like reactions were evaluated.Results The positive group for anaphylactic-anaphylactoid superimposed reaction was determined to be 400 ng·mL-1 DNP-IgE sensitised with 50 ng·mL-1 DNP-BSA and co-administered with C48/80 for 30 min.Additionally,the release of His and β-Hex was promoted by 4,8,and 16 mg·mL-1 of XST in combination with IgE.In the in vivo mouse experiment,5 and 10 mg·kg-1 OVA significantly increased IgE levels in mice.Based on the allergy-like reaction score and the contents of histamine,VEGF-A,and SC5b-9 in the plasma,the 5 mg·kg-1 OVA and 1 mg·kg-1 C48/80 were used to establish a stacked model of allergy-like reaction.There was obvious edema and inflammatory cell infiltration in the ear,lung,and bronchial tissue.XST alone did not sensitize mice,but when combined with OVA-induced allergic reaction,it significantly increased the levels of intra-plasma histamine,VEGF-A,and SC5b-9 compared to the single administration of DNP-IgE/BSA or XST groups(P＜0.05,0.01),showing good consistency with the established allergy-like-allergy stacked model.Conclusion In vitro and in vivo experiments have shown that IgE-mediated allergic reactions and C48/80-induced anaphylactoid reactions produce a superimposed effect,exacerbating the release of allergic mediators and the degree of immune response.The establishment of this model also verified the existence of allergic and anaphylactoid superposition phenomenon of XST.This model can provide a reference for the preclinical safety evaluation and rational use of Chinese medicine injection.
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