Thomazzi Sara M, Moreira Juliana, De Nucci Gilberto, Antunes Edson
Department of Pharmacology, Faculty of Medical Sciences, UNICAMP, P.O. Box 6111, 13084-971 Campinas (SP), Brazil.
Biochem Pharmacol. 2005 Mar 15;69(6):875-82. doi: 10.1016/j.bcp.2004.12.007.
This study was designed to investigate the effects of the 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-pyrimidin-4-ylamine (BAY 41-2272) on formyl-methionyl-leucyl-phenylalanine (fMLP; 10(-7)M)-induced human eosinophil chemotaxis, cyclic guanosine-3',5'-monophosphate (cGMP) and cyclic adenosine-3',5'-monophosphate (cAMP) levels. Human eosinophils were pretreated or not with 3-isobutyl-l-methyl-xanthine (IBMX; 500microM), and then exposed to BAY 41-2272 (0.1-10.0microM) for either short (10min) or prolonged (90min) time periods. Exposition of eosinophils with BAY 41-2272 for either 10min or 90min markedly inhibited the eosinophil chemotaxis, independently of IBMX pretreatment. Inhibition of fMLP-induced eosinophil chemotaxis by BAY 41-2272 (in absence of prior treatment with IBMX) was about of the same irrespective if cells were exposed for 10min or 90min with this compound. In IBMX-pretreated eosinophils, the inhibition of fMLP-induced chemotaxis by BAY 41-2272 in the 10-min exposure protocols was even higher in comparison with the 90-min protocols. Incubation of IBMX-treated eosinophils for 90min with BAY 41-2272 resulted in 2.0-2.5 times higher levels of cGMP and cAMP compared with the 10-min protocols. The BAY 41-2272-induced cGMP increases were abolished by pre-incubation of eosinophils with the soluble guanylate cyclase inhibitor 1H-[1,2,4]-oxidiazolo[4,3-a] quinoxalin-1-one (ODQ). No eosinophil toxicity was observed in any experimental condition, according to 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. Our findings show that inhibitory effects of fMLP-induced human eosinophil chemotaxis by BAY 41-2272 at short-term or prolonged exposition time are accompanied by significant elevations of cGMP and cAMP, but we could not detect a clear correlation between chemotaxis inhibition and elevation of cyclic nucleotide levels.
本研究旨在探讨5-环丙基-2-[1-(2-氟苄基)-1H-吡唑并[3,4-b]吡啶-3-基]-嘧啶-4-胺（BAY 41-2272）对甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP；10⁻⁷M）诱导的人嗜酸性粒细胞趋化性、环磷酸鸟苷（cGMP）和环磷酸腺苷（cAMP）水平的影响。人嗜酸性粒细胞用或不用3-异丁基-1-甲基黄嘌呤（IBMX；500μM）预处理，然后暴露于BAY 41-2272（0.1 - 10.0μM），时间为短时间（10分钟）或长时间（90分钟）。用BAY 41-2272处理嗜酸性粒细胞10分钟或90分钟均显著抑制嗜酸性粒细胞趋化性，与是否用IBMX预处理无关。在未预先用IBMX处理的情况下，BAY 41-2272对fMLP诱导的嗜酸性粒细胞趋化性的抑制作用，无论细胞暴露于该化合物10分钟还是90分钟，效果大致相同。在用IBMX预处理的嗜酸性粒细胞中，与90分钟处理方案相比，在10分钟暴露方案中BAY 41-2272对fMLP诱导的趋化性的抑制作用更强。用BAY 41-2272将用IBMX处理的嗜酸性粒细胞孵育90分钟后，cGMP和cAMP水平比10分钟处理方案高2.0 - 2.5倍。嗜酸性粒细胞与可溶性鸟苷酸环化酶抑制剂1H-[1,2,4] - 恶二唑并[4,3-a]喹喔啉-1-酮（ODQ）预孵育后，BAY 41-2272诱导的cGMP增加被消除。根据3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑（MTT）测定法，在任何实验条件下均未观察到嗜酸性粒细胞毒性。我们的研究结果表明，BAY 41-2272在短期或长期暴露时间对fMLP诱导的人嗜酸性粒细胞趋化性的抑制作用伴随着cGMP和cAMP的显著升高，但我们未检测到趋化性抑制与环核苷酸水平升高之间存在明显相关性。
