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目的:通过比较两种肥大细胞模型RBL-2H3和P815细胞激活后脱颗粒反应的差异,寻找早期、稳定、敏感的肥大细胞脱颗粒体外检测模型,为进一步建立基于细胞水平的过敏原和抗过敏药物的体外筛选模型提供一定的参考。方法:10μg.mL-1C48/80激活细胞15-60min,比色法检测组胺、β-氨基己糖苷酶及类胰蛋白酶等过敏介质的释放,中性红染色法进行形态学观察,流式细胞法检测AnnexinV阳性细胞率。结果:C48/80刺激P815、RBL-2H3细胞15-60min后组胺释放率均明显增加,并且相同刺激条件下,两种细胞模型的组胺释放率基本相当。C48/80刺激P815细胞15min以上,β-氨基己糖苷酶释放率、类胰蛋白酶释放率、Annexin V阳性细胞率和中性红染色脱颗粒细胞率均明显增加;但相同浓度的C48/80需刺激RBL-2H3细胞30-45min以上,上述指标才出现显著增加。在相同的刺激条件下,P815细胞过敏介质释放率、AnnexinV阳性率和脱颗粒率均高于RBL-2H3细胞。结论:同RBL-2H3相比,在相同刺激条件下,P815细胞活化后脱颗粒时间出现更早、程度更高。提示除RBL-2H3细胞,P815细胞也可作为一种早期、稳定、敏感的肥大细胞脱颗粒体外检测模型。
OBJECTIVE: To find an early, stable and sensitive detection model of mast cell degranulation in vitro by comparing the difference of degranulation after activation of RBL-2H3 and P815 cells in two kinds of mast cell models, in order to further establish the cell-level allergen and anti-allergy In vitro drug screening model to provide some reference. Methods: The cells were activated by 10μg.mL-1C48 / 80 for 15-60min. The release of histamine, β-hexosaminidase and tryptase was detected by colorimetric assay. The morphological changes were observed by neutral red staining. AnnexinV positive cells were detected by the cell method. Results: The histamine release rates of P815 and RBL-2H3 cells were significantly increased after 15-60min exposure to C48 / 80, and the histamine release rates of the two cell models were almost the same under the same stimulation conditions. C48 / 80 stimulated P815 cells for more than 15min, the release rate of β-hexosaminidase, tryptase release rate, Annexin V positive cells and neutrophil staining degranulation rate were significantly increased; However, the same concentration of C48 / 80 RBL-2H3 cells stimulated more than 30-45min, the above indicators only a significant increase. Under the same stimulation conditions, the release rate of allergic mediators, the positive rate of AnnexinV and the degranulation rate of P815 cells were higher than that of RBL-2H3 cells. CONCLUSION: Compared with RBL-2H3, P815 cells activated earlier and degranted earlier and to a higher degree under the same stimulation conditions. Tip P815 cells can also be used as an early, stable and sensitive mast cell degranulation test model in vitro except RBL-2H3 cells.