Objective In order to investigate the effect of Osthole,an active ingredient isolated from Chinese herbs Fructus cnidii,on the degranulation of RBL-2H3 cells induced by phorbol.Methods Rat basophilic leukemia (RBL-2H3) cells were cultured in dulbecco's modified eagle medium(DMEM) supplemented with 10％ fetal bovine serum(FBS) and exposed to the varying concentrations of Osthole and phorbol-12-myristate-13-acetate (PMA).The cell viability of the untreated-or treated-cells was determined using cell counting kit-8 (CCK-8) assay;the morphological changes and degranulation of the cells were assessed using phase-contrast microscopy in combination with toluidine blue staining;the activity of β-hexosaminidase of the cells was also determined to estimate the rate of degranulation.Finally,the ultrastructural changes of these cells using transmission electron microscope were examined.Results Compared with untreated control,the cells exposed to 0.5 μmol/L PMA became more rounded,the numbers of intracellular toluidine blue-positive granules and electron-dense granules were significantly decreased in the PMA-treated cells.The activity of β-hexosaminidase of the treated cells was increased as well.Osthole also dose-dependently inhibited the PMA-mediated degranulation in the testing concentrations (1,10,and 50 μmol/L).Our finding suggested that Osthole was a potent inhibitor to block RBL-2H3 cell degranulation.Conclusion Osthole dramatically inhibits non-IgE-mediated degranulation of RBL-2H3 cells,which possibly contributes to its antiallergic activity.%目的 探讨中药活性成分蛇床子素对大鼠RBL-2H3嗜碱性粒细胞脱颗粒的影响.方法 用含10％胎牛血清(FBS)的细胞培养基(Dulbecco's modified eagle medium,DMEM)传代培养RBL-2H3大鼠嗜碱性白血病细胞,并给予不同浓度的蛇床子素和佛波酯(PMA)处理;用CCK-8法测定药物处理与未处理细胞的存活率;油镜结合甲苯胺蓝染色观察细胞形态与脱颗粒变化;测定-氨基己糖苷酶活性估测细胞脱颗粒率;用透射电镜下观察细胞超微结构变化.结果 与空白对照相比,0.5 μmol/L PMA导致细胞明显变圆,胞质内嗜碱性染色(蓝色)的颗粒物减少,-氨基己糖苷酶释放率存在显著增加(P＜0.01);透射电镜也观察到PMA处理细胞内存在较多的空泡,电子致密的粗大颗粒减少,提示PMA能有效刺激RBL-2H3细胞脱颗粒.3个浓度(1,10,50 μmol/L)蛇床子素联合0.5 μmol/L PMA处理细胞,结果显示蛇床子素能剂量依赖地抑制脱颗粒.结论 蛇床子素能明显抑制RBL-2H3细胞非IgE介导脱颗粒反应,可能与其抗过敏活性有关.
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