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J Biol Chem, Vol. 273, Issue 21, 12893-12900, May 22, 1998
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From the Proliferation of vascular smooth
muscle cells (SMC) is a hallmark in the pathogenesis of atherosclerotic
lesions. Mildly oxidized low density lipoproteins (UV-oxLDL), which are
mitogenic to cultured AG-08133A SMC, activate the sphingomyelin
(SM)-ceramide pathway. We report here the following. (i) UV-oxLDL
elicited a biphasic and sustained activation of MBP kinase activity,
phosphorylation and nuclear translocation of p44/42 mitogen-activated
protein kinase (MAPK), and [3H]thymidine
incorporation, which were inhibited by PD-098059, a MAPK kinase
inhibitor. (ii) The use of preconditioned media (from SMC pre-activated
by UV-oxLDL) transferred to native SMC and blocking antibodies against
growth factors suggest that UV-oxLDL-induced activation of MAPK and
[3H]thymidine incorporation seem to be independent of any
autocrine secretion of growth factors. (iii) UV-oxLDL-induced
activation of a neutral sphingomyelinase, SM hydrolysis, ceramide
production, and [3H]thymidine incorporation were
inhibited by two serine-protease inhibitors (serpins), suggesting that
a serpin-sensitive proteolytic pathway is involved in the activation of
the SM-ceramide signaling pathway. (iv) UV-oxLDL-induced MAPK
activation and [3H]thymidine incorporation were mimicked
by ceramide generated in the plasma membrane by bacterial
sphingomyelinase treatment or by addition of the permeant
C2-ceramide. Serpins did not inhibit the MAPK activation
and [3H]thymidine incorporation induced by
C2-ceramide, indicating that activation of the MAPK and
[3H]thymidine incorporation is subsequent to the
stimulation of the SM-ceramide pathway. Taken together, these data
suggest that mitogenic concentrations of UV-oxLDL are able to stimulate
the SM-ceramide pathway through a protease-dependent
mechanism and activate p44/42 MAPK, leading to proliferation of
vascular SMC.
Laboratory of Biochemistry, INSERM U-466,
Institut Louis Bugnard, CHU Rangueil, 1 Avenue Jean Poulhes, 31403 Toulouse Cedex 4, France and INSERM
CJF-9503, Centre Claudius Regaud, 20 Rue du Pont Saint-Pierre,
31052 Toulouse Cedex, France
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