Vascular smooth muscle cells on Matrigel as a model for LPS-induced hypocontractility and NO formation.
Treatment of vascular tissue with low levels of lipopolysaccharide (LPS) induces nitric oxide synthase (NOS) activity and diminishes vascular contractility. However, in cultured vascular smooth muscle cells (VSMC), very high doses of LPS or the combination of LPS with cytokines are required for the induction of nitric oxide (NO) formation. The aims of this study were to establish a cell model to investigate LPS-induced hypocontractility and NO production and to test the hypothesis that responses of VSMC to LPS are differentiation regulated. We used Matrigel basement membrane matrix to maintain VSMC differentiation and found that VSMC cultured on Matrigel retained significant contractility in response to KCl stimulation. Incubation of VSMC with low levels of LPS(1-100 ng/ml) induced NOS mRNA and protein, induced NO production, and decreased cell contractility in a time- and dose-dependent fashion. The NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) partially restored LPS-treated VSMC contractility, whereas L-arginine reversed the contractility-restoring effect of L-NAME. These results suggest that VSMC grown on Matrigel are a useful experimental model for investigations into signal transduction mechanisms responsible for LPS-induced vascular hypocontractility.
Li
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Septic Shock Research Program, Naval Medical Research Institute, Bethesda, Maryland 20889-5607, USA.
Fan
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SX
McKenna
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TM
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen; D004338:Drug Combinations; D004790:Enzyme Induction; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007797:Laminin; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014661:Vasoconstriction / Q000187:drug effects*
D004338:Drug Combinations; D004791:Enzyme Inhibitors; D007797:Laminin; D008070:Lipopolysaccharides; D011509:Proteoglycans; D012333:RNA, Messenger; C059485:matrigel; D009569:Nitric Oxide; D003094:Collagen; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester