Teshima R, Ikebuchi H, Terao T, Nakanishi M
National Institute of Hygienic Sciences, Tokyo, Japan.
Chem Pharm Bull (Tokyo). 1991 Mar;39(3):747-51. doi: 10.1248/cpb.39.747.
We examined the effect of staurosporine on cytosolic calcium response in rat basophilic leukemia (RBL-2H3) cells using fura-2 as a fluorescent indicator of calcium ion. Staurosporine at a dose of 30 nM inhibited antigen-stimulated Ca2(+)-influx into the cells from the extracellular environment. In contrast, the drug at this concentration inhibited neither the mobilization of Ca2+ from intracellular stores nor inositol 1,4,5-trisphosphate (IP3) formation. At a high concentration (300 nM), however, staurosporine completely inhibited the cytosolic calcium responses as well as IP3 formation. These results indicate that staurosporine, if used at an appropriate concentration, can be used to discriminate Ca2(+)-influx from extracellular environment from mobilization of the ion from intracellular stores. These results also suggest that protein kinases, possibly protein kinase C, are involved in the calcium influx of RBL-2H3 cells from the extracellular environment. Serotonin release was strongly inhibited by the drug at 30 nM staurosporine. Since the inhibition of serotonin release and suppression of cytosolic calcium increase in response to the antigen were in parallel, we concluded that the inhibition of serotonin release from RBL-2H3 cells caused by the drug was elicited by the suppression of Ca2(+)-influx into the cells.
我们使用fura-2作为钙离子的荧光指示剂，研究了星形孢菌素对大鼠嗜碱性白血病（RBL-2H3）细胞胞质钙反应的影响。30 nM剂量的星形孢菌素抑制了抗原刺激的Ca2+从细胞外环境流入细胞。相比之下，该浓度的药物既不抑制细胞内钙库中Ca2+的释放，也不抑制肌醇1,4,5-三磷酸（IP3）的形成。然而，在高浓度（300 nM）时，星形孢菌素完全抑制了胞质钙反应以及IP3的形成。这些结果表明，星形孢菌素如果以适当的浓度使用，可用于区分细胞外环境中的Ca2+流入与细胞内钙库中离子的释放。这些结果还表明，蛋白激酶，可能是蛋白激酶C，参与了RBL-2H3细胞从细胞外环境的钙流入。30 nM星形孢菌素强烈抑制了5-羟色胺的释放。由于5-羟色胺释放的抑制和抗原刺激引起的胞质钙增加的抑制是平行的，我们得出结论，该药物引起的RBL-2H3细胞5-羟色胺释放的抑制是由Ca2+流入细胞的抑制引起的。
