羧胺三唑对RBL-2H3细胞活化脱颗粒的影响

Effects of carboxyamidotriazole on the activation and degranulation of RBL-2H3 cells

目的研究羧胺三唑(CAI)对RBL-2H3肥大细胞增殖、凋亡及活化脱颗粒的影响,探索CAI的抗感染作用机制。方法以C48/80诱导RBL-2H3细胞活化脱颗粒模型,中性红染色法观察细胞脱颗粒的形态学,分别用ELISA法和底物显色法检测细胞培养上清中组胺和β-氨基己糖苷酶的释放水平,CCK-8法测定细胞活力,Hoechst 33342荧光染色法检测细胞凋亡。结果与对照组相比,10、20和40μmol/L CAI能够不同程度抑制C48/80诱导的RBL-2H3细胞脱颗粒反应,20和40μmol/L CAI能够降低C48/80诱导的组胺释放(P<0.01),40μmol/L CAI能够降低β-氨基己糖苷酶的释放(P<0.01)。另外,所用各浓度的CAI对细胞增殖和凋亡均无明显影响。结论 CAI能有效抑制RBL-2H3肥大细胞的活化脱颗粒,此作用并不是通过细胞毒发挥作用的。CAI可能部分通过下调肥大细胞的功能活化,发挥其抗感染作用。

Objective To explore the anti-infection mechanism of carboxyamidotriazole（ CAI） through studying the effects of CAI on the proliferation,apoptosis and degranulation of RBL-2H3 mass cells. Methods Compound 48/80（ C48/80） was used to induce the model of activation and degranulation in RBL-2H3 cells. The morphological change of cell degranulation was observed by neutral red staining. The release levels of histamine and β-hexosaminidase were measured by ELISA method and chromogenic assay,respectively. The cell activity was determined by CCK-8 method. And cell apoptosis was detected by Hoechst 33342 fluorescent staining. Results Compared with the control group,10,20,40 μmol/L CAI inhibited C48/80-induced degranulation of RBL-2H3 cells in different degrees. CAI（ 20,40 μmol/L） reduced the histamine release（ P〈0. 01）,and CAI（ 40 μmol/L） decreased theβ-hexosaminidase release（ P〈0. 01）. In addition,the viability and apoptosis of RBL-2H3 cells were not affected at the concentrations of CAI used. Conclusions CAI can effectively inhibit the activation and degranulation of RBL-2H3 mast cells,and this effect is not through cytotoxicity. The anti-infection effect of CAI may partially due to the down-regulation of mast cell activity.