Establishment of In-vitro Model of Mast Cell Degranulation with Three Kinds of Cell Lines and Its Application

Objective To investigate the difference between C48/80-induced degranulation in Ku812,RBL-2H3,and P815 cells,and to find an early,stable and sensitive in-vitro mast cell degranulation model for the detection of allergic and anaphylactoid reaction induced by several traditional Chinese medicinal injections(TCMI).Methods Cells were activated with 10 μg/mL of C48/80 for 0-60 min.Cell degranulation was evaluated by detecting the release of histamine with liquid chromatography-mass chromatography(LC-MS),examining hexosaminidase with colorimetric assays,and observing the morphological changes with neutral red staining.Results After treatment with C48/80,the histamine release rate,hexosaminidase release rate and cell degranulation rate were markedly increased in Ku812,RBL-2H3 and P815 cells(P0.05),and the degranulation occurred earlier and was more obvious in the activated P815 cells(P0.05).And then the in-vitro detection of the sensitization of several TCMI was performed on P815 cells.The results showed that Qingkailing Injection,Xuesaitong Injection,Shuanghuanglian Injection,Zhonjiefeng Injection and Xiangdan Injection induced obvious release of histamine in P815 cells.Conclusion Compared with Ku812 and RBL-2H3 cells,P815 cells are suitable for the early,stable and sensitive in-vitro mast cell degranulation model,and the detection results of allergic anaphylactoid reaction induced by several common TCMI show certain accordance with the clinical reports.P815 cells can be expected to evaluate the allergic and anaphylactoid reaction caused by TCMI.