Adult cardiac fibroblasts null for sphingosine kinase-1 exhibit growth dysregulation and an enhanced proinflammatory response.
Cardiac fibroblasts are critical for the maintenance of extracellular matrix deposition and turnover in the normal heart and are key mediators of inflammatory and fibrotic myocardial remodeling in the injured and failing heart. Sphingosine kinase (SphK) activation is a well-recognized determinant of cell fate in cardiac myocytes and other cells, but SphK responses have not previously been studied in cardiac fibroblasts. Initially we found that total SphK activity is over 10-fold higher in cardiac fibroblasts than in adult mouse cardiac myocytes. SphK is composed of two major isoforms, SphK-1 and SphK-2. In fibroblasts isolated from SphK-1 knockout mice, SphK activity was greatly reduced indicating that SphK-1 is the major isoform expressed in these cells. To determine whether SphK regulates cell proliferation and the proinflammatory protein inducible nitric oxide synthase (iNOS), we exposed cultured cardiac fibroblasts to the cytokine interleukin-1beta (IL-1beta) and/or hypoxia. Both hypoxia and IL-1beta alone and in combination enhanced fibroblast SphK activity. In wild-type fibroblasts, hypoxia induced proliferation, but in SphK-1 null fibroblasts this response was blunted even in the presence of serum. In contrast, we found that iNOS expression and NO production were enhanced in SphK-1 null fibroblasts during hypoxia. In wild-type fibroblasts, IL-1beta was only a weak inducer of iNOS and of NO accumulation and hypoxia alone had no significant effect on iNOS activation. However, IL-1beta in combination with hypoxia extensively stimulated iNOS and NO production, and this stimulation was enhanced in SphK-1 null fibroblasts. We conclude that activation of endogenous SphK-1 serves a dual regulatory function: it is required for optimal cardiac fibroblast proliferation but is a negative modulator of proinflammatory responses during hypoxia.
Kacimi
Rachid
R
Cardiology Research, Cardiology Section (111C5), VA Medical Center, San Francisco, CA 94121, USA.
Vessey
Donald A
DA
Honbo
Norman
N
Karliner
Joel S
JS
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D000860:Hypoxia; D053583:Interleukin-1beta / Q000378:metabolism* / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009206:Myocardium / Q000166:cytology*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D000099157:Sphingosine Kinase
D053583:Interleukin-1beta; D007527:Isoenzymes; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D017853:Phosphotransferases (Alcohol Group Acceptor); D000099157:Sphingosine Kinase