NGF exhibits a pro-apoptotic activity for human vascular smooth muscle cells that is inhibited by TGFbeta1.
Apoptosis of vascular smooth muscle cells (SMCs) has been described in culture and also during remodelling of the artery following injury. However, the mediators that regulate apoptosis in SMCs are unknown. Because neurotrophins, a family of related polypeptide growth factors, including nerve growth factor (NGF) and its cognate receptor TrkA have been shown to be strongly expressed in atherosclerotic lesions, the present study was undertaken to evaluate in vitro, the activity of NGF with regard to apoptosis of confluent cultures of human aortic SMCs. We report here that NGF induced apoptosis of SMCs in a dose-dependent manner. This effect was detected from the concentration of 1 ng/ml and reached a maximum at 100 ng/ml. The concentration that induced a half-maximum effect was 8.8 ng/ml. The pro-apoptotic activity of NGF was time dependent and was significant after 3 h of incubation. The pro-apoptotic activity of NGF was blocked in a dose-dependent manner by K-252a, an inhibitor of TrkA tyrosine phosphorylation, suggesting that a NGF/TrkA signal transduction pathway could activate apoptotic cell death programs in human SMCs. Significantly, NGF-induced apoptosis was inhibited by wortmannin and PD 98059, showing that both PI3 kinase and MEK kinase were involved. At a NGF concentration that strongly induced apoptosis (100 ng/ml), TGFbeta1 which has been identified several times as a protective factor, dose dependently inhibited the pro-apoptotic effect of NGF. The IC50 value was 1.5 ng/ml. These results indicate that, at least in vitro, TGFbeta1 can inhibit the pro-apoptotic activity of NGF for SMCs therefore suggesting that TGFbeta1 has the capacity to diminish the deleterious consequences of an excitotoxic or ischemic injury that might occur during atherogenesis or following angioplasty.
Bono
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Haemobiology Research Department, Sanofi Recherche, Toulouse, France.
Lamarche
I
I
Herbert
J M
JM
D000730:Androstadienes / Q000494:pharmacology; D001011:Aorta; D017209:Apoptosis / Q000187:drug effects*; D002227:Carbazoles / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D026121:Indole Alkaloids; D007700:Kinetics; D048728:MAP Kinase Kinase Kinase 1; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D009414:Nerve Growth Factors / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000502:physiology; D020794:Receptor Protein-Tyrosine Kinases / Q000502:physiology; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D000077191:Wortmannin
D000730:Androstadienes; D002227:Carbazoles; D004791:Enzyme Inhibitors; D005419:Flavonoids; D026121:Indole Alkaloids; D048728:MAP Kinase Kinase Kinase 1; D009414:Nerve Growth Factors; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D020794:Receptor Protein-Tyrosine Kinases; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor; D016212:Transforming Growth Factor beta; D000077191:Wortmannin; C049985:staurosporine aglycone; C482576:MAP3K1 protein, human; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one