TY - JOUR
T1 - Induction and activity of nitric oxide synthase in cultured human intestinal epithelial monolayers
AU - Salzman, Andrew L.
AU - Denenberg, Alvin G.
AU - Ueta, Ikuya
AU - O'Connor, Michael
AU - Linn, Stephen C.
AU - Szabó, Csaba
PY - 1996
Y1 - 1996
N2 - We have examined the induction and activity of inducible nitric oxide (NO) synthase (iNOS) in monolayers of DLD-1 cells, an epithelial cell line derived from a human intestinal adenocarcinoma. Induction of iNOS transcription by a combination of the cytokines interferon-γ and IL-1β was inhibited by genistein, pyrrolidine dithiocarbamate, or dexamethasone and unaffected by pretreatment with ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'- tetraacetic acid, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), or the isoflavone daidzein. iNOS activity and NO synthesis were inhibited by nitro-L-arginine methyl ester, N(G)-mono-methyl-L-arginine, S- methyl-isothiourea sulfate, or amino-ethyl-isothiourea, but not by dexamethasone. NO synthesis was potently inhibited by N-α-p-tosyl-lysine chloromethyl ketone and hypoxia. In the absence of cytokines, no iNOS induction was observed with oxidant stress (H2O2), growth factors (bFGF, EGF), hypoxia, or hypoxia reoxygenation. We conclude that in this model of the human intestinal epithelium 1) cytokine-mediated induction of iNOS is Ca2+ independent, weakly steroid sensitive, and may involve the activation of nuclear factor-κB and a tyrosine kinase, and 2) iNOS activity is Ca2+- independent and inhibited by hypoxia, N(G)-substituted L-arginine analogues, and isothioureas.
AB - We have examined the induction and activity of inducible nitric oxide (NO) synthase (iNOS) in monolayers of DLD-1 cells, an epithelial cell line derived from a human intestinal adenocarcinoma. Induction of iNOS transcription by a combination of the cytokines interferon-γ and IL-1β was inhibited by genistein, pyrrolidine dithiocarbamate, or dexamethasone and unaffected by pretreatment with ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'- tetraacetic acid, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), or the isoflavone daidzein. iNOS activity and NO synthesis were inhibited by nitro-L-arginine methyl ester, N(G)-mono-methyl-L-arginine, S- methyl-isothiourea sulfate, or amino-ethyl-isothiourea, but not by dexamethasone. NO synthesis was potently inhibited by N-α-p-tosyl-lysine chloromethyl ketone and hypoxia. In the absence of cytokines, no iNOS induction was observed with oxidant stress (H2O2), growth factors (bFGF, EGF), hypoxia, or hypoxia reoxygenation. We conclude that in this model of the human intestinal epithelium 1) cytokine-mediated induction of iNOS is Ca2+ independent, weakly steroid sensitive, and may involve the activation of nuclear factor-κB and a tyrosine kinase, and 2) iNOS activity is Ca2+- independent and inhibited by hypoxia, N(G)-substituted L-arginine analogues, and isothioureas.
KW - calcium
KW - isothiourea
KW - nuclear factor-κB
KW - steroids
KW - tyrosine kinase
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U2 - 10.1152/ajpgi.1996.270.4.g565
DO - 10.1152/ajpgi.1996.270.4.g565
M3 - Article
C2 - 8928785
AN - SCOPUS:0029881475
SN - 0193-1857
VL - 270
SP - G565-G573
JO - American Journal of Physiology - Gastrointestinal and Liver Physiology
JF - American Journal of Physiology - Gastrointestinal and Liver Physiology
IS - 4 33-4
ER -