Experimental study on the allergenicity of active ingredients in injectable Shuanghuangli-an based on molecular docking technology
Objective Screening for the presence of allergenic components in Shuanghuanglian injection and a preliminary inves-tigation of the mechanism of action.Methods With the target protein of Mas-related G protein-coupled receptor X2(MRG-PRX2)as the target protein,molecular docking with the active substance in injectable Shuanghuanglian as the ligand was carried out by AutoDock and Pymol software to preliminarily screen for class allergenic components,and the optimal RBL-2H3 cell ex-perimental conditions(IC50)were examined,the initially selected components of the active ingredients of Shuanghuanglian injec-tion were incubated with RBL-2H3 cells,and the release rate of histamine and β-aminohexosidase was detected by double-antibody sandwich ELISA with substrate assay,to further screen the class-causing periplasmic components,and to observe their effects on the intracellular complement level and the degree of cell degranulation in RBL-2H3 cells,which were finally verified by a rat isolated ileum experiment for verification.Results His and β-Hex release were significantly elevated in RBL-2H3 cells after administration of Shuanghuanglian injection(28.25 mg/L),bate-sitosterol,(0.95 mg/L),bate-carotene(4.10 mg/L),baicalein(1.90 mg/L),methyl caffeate(3.20 mg/L),ferulic acid(2.29 mg/L)and chlorogenic acid(1.51 mg/L),respectively(P＜0.01,0.05).Total complement level CH50 and C3a and C5a levels were significantly elevated(P＜0.05)with degranulation in rats after administration.After isolated ileum test β-sitosterol,β-carotene,baicalein,methyl caffeate,ferulic acid and chlorogenic acid increased ileal tone in rats(P＜0.01,0.05).Conclusion The anaphylactoid reaction induced by Shuanghuanglian injection may be related to β-sitosterol,β-carotene,baicalein,methyl caffeate,ferulic acid and chloro-genic acid,and the mechanism of action of the induced anaphylactoid reaction may be related to the level of complement.
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