RBL-2H3和P815细胞用于建立体外肥大细胞脱颗粒模型的比较研究(英文)

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...

Comparison of RBL-2H3 and P815 Cell Lines for Establishing in vitro Model of Mast Cell Degranulation

AIM： To investigated the difference between C48/80-induced degranulation in RBL-2H3 and that in P815 cells, and to find an early, stable and sensitive in vitro mast cell degranulation model. All findings might provide reference for establishing in vitro models to screen allergens and testing anti-allergic agents. METHODS： Cells were treated with 10 μg·mL^-1 C48/80 for fifteen to sixty minutes. Cell degranulation was evaluated by detecting the release of histamine,β-hexosaminidase and tryptase by colorimetric assays, observing the morphological changes by neutral red staining, and detecting the ratio of Annexin V positive cell by flow cytometry. RESULTS： After treatment with C48/80, the histamine release in RBL-2H3 and P815 cells was significantly increased （P 〈 0.05）, and the ratio of histamine release in both cells was almost equivalent, β-hexosaminidase release, tryptase release, the percentage of Annxin V positive cells and the degranulation ratio were markedly elevated in P815 cells treated with C48/80 for more than 15 min, but the same concentration of C48/80 induced similar changes in RBL-2H3 cells only when the cells were treated for more than 30 to 45 min. Upon the same stimulating conditions, allergic mediators release, the percentage of Annxin V positive cells and the degranulation ratio in P815 cells were much higher than those in RBL-2H3 cells. CONCLUSION： Compared with the RBL-2H3 cells, cell degranulation of P815 cells occurred faster and more severe than that of RBL-2H3 under the s ame stimulating conditions. These data sustain our contention that P815 cells might be utilized as a potential early, fast and sensitive in vitro model for screening suspicious allergens and searching for anti-allergic agents.