Sphingosine-1-phosphate stimulates smooth muscle cell migration through Gαi- and PI3-kinase-dependent P38MAPK activation

被引:28
作者
Fegley, AJ
Tanski, WJ
Roztocil, E
Davies, MG
机构
[1] Univ Rochester, Med Ctr, Vasc Biol & Therapeut Program, Ctr Vasc Dis,Dept Surg,Div Vasc Surg, Rochester, NY 14642 USA
[2] Univ Rochester, Cardiovasc Res Ctr, Rochester, NY 14642 USA
关键词
sphingosine-1-phosphate; p38 MAP kinase; smooth muscle cells; signal transduction; migration; G-protein signaling; PI3-kinase;
D O I
10.1016/S0022-4804(03)00120-3
中图分类号
R61 [外科手术学];
学科分类号
摘要
Background. Sphingosine-1-phosphate (S-1-P) is an extracellular mediator released in response to vessel injury. S-1-P binds to G-protein-coupled receptors, which can be Galpha(i)-, Galpha(q)-, or G(12/13)-linked. This study examines the role of p38 mitogen-activated protein kinase (p38(MAPK)) in vascular smooth muscle cell migration after stimulation with S-1-P, and pathways leading to p38(MAPK) activation. S-1-P has previously been shown to stimulate migration of vascular smooth muscle cells (VSMCs) in vitro through ERK1/2 and G(i). We hypothesized that S-1-P-induced VSMC migration is also dependent on p38(MAPK) activation through a G(i)-coupled extracellular receptor and phosphoinositide 3-kinase (PI3-K). Methods. VSMCs, were cultured in vitro. A linear wound assay was performed in the presence of S-l-P and inhibitors of p38(MAPK) (SB203580) or epidermal growth factor (EGF) receptor kinase (AG1478). Chemotaxis stimulated by S-I-P was also assayed in a modified Boyden chamber with and without SB203580 pretreatment. Western blotting was performed to examine p38(MAPK) activation in response to S-1-P with and without SB203580, AG1478, or inhibitors of Gi (pertussis toxin), PI3-K (Wortmannin and LY294002), or MEK1 (PD98059). Western blotting and immunoprecipitation for targets of p38(MAPK) (MAPKAP kinase-2) and PI3-K (Akt) were also performed. Results. S-1-P stimulated migration of VSMCs in both wound and Boyden transwell assays. This migration was inhibited by SB203580 to the level of control, whereas AG478 had no effect. S-1-P stimulated activation of p38(MAPK) that peaked at 10 min, as well as activation of MAPKAP kinase-2. Activation of p38(MAPK) was significantly inhibited by SB203580, pertussis toxin, Wortmannin, and LY294002, but not by PD98059 or AG1478; MAPKAP kinase-2 activation was inhibited by SB203580. Akt was activated by S-1-P at 3 to 5 min; this response was inhibited by Wortmannin and LY294002, but not by SB203580 or pertussis toxin. Conclusions. S-1-P induced VSMC migration through a G(i)-linked and a PI3-K coupled, p38(MAPK)-dependent process. PI3-K appears to function upstream of p38(MAPK), but was not G(i)-dependent. S-1-P-stimulated activation of p38(MAPK) does not signal via transactivation of the EGF receptor. Understanding signal transduction will allow targeted molecular interventions to treat the response of a vessel to injury. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:32 / 41
页数:10
相关论文
共 37 条
[1]   PD-098059 IS A SPECIFIC INHIBITOR OF THE ACTIVATION OF MITOGEN-ACTIVATED PROTEIN-KINASE KINASE IN-VITRO AND IN-VIVO [J].
ALESSI, DR ;
CUENDA, A ;
COHEN, P ;
DUDLEY, DT ;
SALTIEL, AR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (46) :27489-27494
[2]   SB-203580 IS A SPECIFIC INHIBITOR OF A MAP KINASE HOMOLOG WHICH IS STIMULATED BY CELLULAR STRESSES AND INTERLEUKIN-1 [J].
CUENDA, A ;
ROUSE, J ;
DOZA, YN ;
MEIER, R ;
COHEN, P ;
GALLAGHER, TF ;
YOUNG, PR ;
LEE, JC .
FEBS LETTERS, 1995, 364 (02) :229-233
[3]   PATHOBIOLOGY OF INTIMAL HYPERPLASIA [J].
DAVIES, MG ;
HAGEN, PO .
BRITISH JOURNAL OF SURGERY, 1994, 81 (09) :1254-1269
[4]   Specificity and mechanism of action of some commonly used protein kinase inhibitors [J].
Davies, SP ;
Reddy, H ;
Caivano, M ;
Cohen, P .
BIOCHEMICAL JOURNAL, 2000, 351 (351) :95-105
[5]   Phosphatidylinositol 3-kinase is required for insulin-like growth Factor-I-Induced vascular smooth muscle cell proliferation and migration [J].
Duan, C ;
Bauchat, JR ;
Hsieh, T .
CIRCULATION RESEARCH, 2000, 86 (01) :15-23
[6]   Calcium-dependent epidermal growth factor receptor transactivation mediates the angiotensin II-induced mitogen-activated protein kinase activation in vascular smooth muscle cells [J].
Eguchi, S ;
Numaguchi, K ;
Iwasaki, H ;
Matsumoto, T ;
Yamakawa, T ;
Utsunomiya, H ;
Motley, ED ;
Kawakatsu, H ;
Owada, KM ;
Hirata, Y ;
Marumo, F ;
Inagami, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (15) :8890-8896
[7]   Sphingosine 1-phosphate released from platelets during clotting accounts for the potent endothelial cell chemotactic activity of blood serum and provides a novel link between hemostasis and angiogenesis [J].
English, D ;
Welch, Z ;
Kovala, AT ;
Harvey, K ;
Volpert, OV ;
Brindley, DN ;
Garcia, JGN .
FASEB JOURNAL, 2000, 14 (14) :2255-2265
[8]   Platelet-released phospholipids link haemostasis and angiogenesis [J].
English, D ;
Garcia, JGN ;
Brindley, DN .
CARDIOVASCULAR RESEARCH, 2001, 49 (03) :588-599
[9]   Signal Transduction in smooth muscle - Invited review: Focal adhesion and small heat shock proteins in the regulation of actin remodeling and contractility in smooth muscle [J].
Gerthoffer, WT ;
Gunst, SJ .
JOURNAL OF APPLIED PHYSIOLOGY, 2001, 91 (02) :963-972
[10]   Diversity of cellular receptors and functions for the lysophospholipid growth factors lysophosphatidic acid and sphingosine 1-phosphate [J].
Goetzl, EJ ;
An, SZ .
FASEB JOURNAL, 1998, 12 (15) :1589-1598