Abstract
We now evaluated the anti-inflammatory mechanisms of andrographolide on complement 5a (C5a)-induced macrophage recruitment in vitro. Andrographolide concentration dependently inhibited cell migration toward C5a with an IC50 of 5.6+/-0.7 microM. With relatively specific kinase inhibitors (PD98059, SB203580, SP600125, wortmannin and LY294002, respectively) the results showed that extracellular signal-regulated kinase1/2 (ERK1/2), p38 mitogen-activated protein kinase (p38 MAPK) and phosphatidylinositol-3-kinase (PI3K) were necessary for C5a-induced migration, whereas c-Jun N-terminal kinase (JNK) was nonessential. Andrographolide significantly attenuated C5a-stimulated phosphorylation of ERK1/2, and of its upstream activator, MAP kinase-ERK kinase (MEK1/2). C5a-activated ERK1/2 phosphorylation was 86+/-9% inhibited by 30 microM andrographolide. Under the same conditions, however, andrographolide failed to affect C5a-stimulated p38 MAPK and JNK phosphorylation. Andrographolide also strongly abolished C5a-stimulated Akt phosphorylation, a downstream target protein for PI3K. These results indicate that inhibition of cell migration by interfering with ERK1/2 and PI3K/Akt signal pathways may contribute to the anti-inflammatory activity of andrographolide.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Androstadienes / pharmacology
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Animals
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Anthracenes / pharmacology
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Cell Line
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Cell Movement / drug effects*
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Cell Survival / drug effects
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Chemokine CCL4
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Chemotaxis / drug effects
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Chromones / pharmacology
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Complement C5a / pharmacology
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Diterpenes / pharmacology*
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Dose-Response Relationship, Drug
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Enzyme Inhibitors / pharmacology
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Flavonoids / pharmacology
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Imidazoles / pharmacology
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JNK Mitogen-Activated Protein Kinases / metabolism
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MAP Kinase Kinase 1 / metabolism
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MAP Kinase Kinase 2 / metabolism
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MAP Kinase Kinase 4
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Macrophage Inflammatory Proteins / pharmacology
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Mitogen-Activated Protein Kinase 1 / metabolism*
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Mitogen-Activated Protein Kinase 3 / metabolism*
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Mitogen-Activated Protein Kinase Kinases / metabolism
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Morpholines / pharmacology
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Phosphoinositide-3 Kinase Inhibitors
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Phosphorylation / drug effects
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Protein Serine-Threonine Kinases / metabolism*
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Proto-Oncogene Proteins / metabolism*
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Proto-Oncogene Proteins c-akt
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Pyridines / pharmacology
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Wortmannin
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p38 Mitogen-Activated Protein Kinases / metabolism
Substances
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Androstadienes
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Anthracenes
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Chemokine CCL4
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Chromones
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Diterpenes
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Enzyme Inhibitors
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Flavonoids
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Imidazoles
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Macrophage Inflammatory Proteins
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Morpholines
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Phosphoinositide-3 Kinase Inhibitors
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Proto-Oncogene Proteins
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Pyridines
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pyrazolanthrone
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2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
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andrographolide
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Complement C5a
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Protein Serine-Threonine Kinases
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Proto-Oncogene Proteins c-akt
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JNK Mitogen-Activated Protein Kinases
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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p38 Mitogen-Activated Protein Kinases
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MAP Kinase Kinase 1
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MAP Kinase Kinase 2
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MAP Kinase Kinase 4
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Map2k1 protein, mouse
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Map2k2 protein, mouse
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Mitogen-Activated Protein Kinase Kinases
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SB 203580
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2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
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Wortmannin