Nitric oxide production stimulated by the basic fibroblast growth factor requires the synthesis of ceramide

被引:11
作者
Arena, S
Pattarozzi, A
Thellung, S
Villa, V
Corsaro, A
Massa, A
Diana, F
Spoto, G
Forcella, S
Damonte, G
Filocamo, M
Benatti, U
Schettini, G
Florio, T
机构
[1] CBA, Unita Neurosci, Adv Biotechnol Ctr, Natl Inst Canc Res,IST, I-16132 Genoa, Italy
[2] Univ Genoa, Dept Oncol Biol & Genet, Pharmacol Sect, Genoa, Italy
[3] Univ G DAnnunzio, Dept Biomed Sci, Pharmacol Sect, Chieti, Italy
[4] Univ G DAnnunzio, Dept Appl Sci Oral & Dent Dis, Chieti, Italy
[5] Univ Genoa, Dept Expt Med, Genoa, Italy
[6] Ist Giannina Gaslini, Diagnost Lab Pre & Postnatal Metab Disorders, Genoa, Italy
来源
CELL SIGNALING, TRANSCRIPTION, AND TRANSLATION AS THERAPEUTIC TARGETS | 2002年 / 973卷
关键词
bFGF; nitric oxide; ceramide;
D O I
10.1111/j.1749-6632.2002.tb04614.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nitric oxide (NO) is an intracellular and intercellular mediator involved in the modulation of many physiologic and pathologic processes including the regulation of neoangiogenesis. We analyzed the effects of basic fibroblast growth factor (bFGF) on NO production in CHO-K1 cells and the intracellular mechanisms involved. bFGF induces NO production through activation of the endothelial NO synthase (eNOS), causing a subsequent increase in cGMP levels. In most systems, eNOS activation is a Ca2+-calmodulin-dependent process. In CHO-K1 cells, NO production by bFGF is Ca2+ and MAP kinase independent, because it was not reverted by pretreatment with intracellular Ca2+ chelators or MEK inhibitors. Translocation of the eNOS from the plasma membrane, where it is bound to caveolin 1, to the cytosol is the crucial step in the synthesis of NO. We demonstrate that the cytosolic translocation of eNOS is caused by increased synthesis of ceramide dependent by the bFGF activation of sphingomyelinase. Indeed, in the presence of the sphingomyelinase inhibitors D609 or desipramine, bFGF-dependent NO production is abrogated. To support this evidence we evaluated ceramide concentration using HPLC-electrospray ionization-mass spectrometry in controls and in bFGF-treated cells: after bFGF stimulation, a substantial increase in ceramide levels was observed. These data were further confirmed by the lack of NO production in response to fibroblast growth factor in fibroblasts derived from Niemann Pick patients who genetically lack the enzyme sphingomyelinase. In conclusion, ceramide in CHO-K1 cells is responsible for a novel Ca2+/Calmodulin-independent mechanism for eNOS activation after fibroblast growth factor stimulation.
引用
收藏
页码:94 / 104
页数:11
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