| Anaphylactoid reaction is one of the main adverse reactions of Traditional Chinese Medicine Injections(TCMIs)in clinic.Therefore,to ensure safe and efficient applications of TCMIs,it is important to identify allergens in TCMIs and perform mechanistic studies.Current study was designed to screen and verify the allergen responsible for anaphylactoid reactions through in vivo and in vitro assays.Besides,the metabolomic profiles were performed to explore the differential metabolites and elucidate the mechanisms that may be associated with anaphylactoid reactions after the allergen administration.Results from this work would provide valuable scientific information for further re-evaluation of TCMIs.The main studies were shown as below:(1)The degranulation model of RBL-2H3 cell with the endpoint ofβ-hexosaminidase release rate was established to screen eight TCMIs.One of these injections was observed to induce obvious degranulation in RBL-2H3 cells.Further screening showed that Ginsenoside Rd,one of the five main components of this injection,could significantly increase β-hexosaminidase release and IL-4 level in a dose-dependent manner in RBL-2H3 cells.Preliminary studies indicated Rd could affect the p38 MAPK,ERK1/2 and JNK phosphorylation levels in the MAPK pathway,suggesting this pathway may play an important role in Rd induced mast cell degranulation.In addition,a mice cutaneous vascular permeability assay was applied.The results showed that Rd significantly increased the vascular permeability in a dose-dependent manner,which was consistent with those of in vitro.(2)Moreover,Rd significantly elevated the histamine level in plasma after tail intravenous injection in mice.Besides,a comprehensive metabolomics approach was established to investigate the effects of Rd-induced anaphylactoid reaction in mice using ultra performance liquid chromatograph quadrupole time-of-flight-mass spectrometry(UPLC-QTOF/MS).The samples of 10 min,30 min,and 2 h after Rd injection were collected and used for metabolomics analysis.Partial Least Squares-Discriminant Analysis(PLS-DA)and Orthogonal Partial Least Squares-Diserminant Analysis(OPLS-DA)showed that the the samples of Rd groups were clearly separated from samples of control groups at the three time-points.The metabolomic profiles were changed in a time-dependent manner,and the samples of Rd group at 2 h displayed a trend towards to control group.Furthermore,the metabolites analysis showed that Rd-induced anaphylactoid reactions may be closely related to energy metabolism,signal transduction and inflammation or immune response. |
