NO and TNF-alpha released from activated macrophages stabilize HIF-1alpha in resting tubular LLC-PK1 cells.
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NO and TNF-alpha released from activated macrophages stabilize HIF-1alpha in resting tubular LLC-PK1 cells. / Zhou, Jie; Fandrey, Joachim; Schümann, Jens; Tiegs, Gisa; Brüne, Bernhard.
in: AM J PHYSIOL-CELL PH, Jahrgang 284, Nr. 2, 2, 2003, S. 439-446.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - NO and TNF-alpha released from activated macrophages stabilize HIF-1alpha in resting tubular LLC-PK1 cells.
AU - Zhou, Jie
AU - Fandrey, Joachim
AU - Schümann, Jens
AU - Tiegs, Gisa
AU - Brüne, Bernhard
PY - 2003
Y1 - 2003
N2 - Hypoxic/ischemic conditions provoke activation of the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 is composed of HIF-1alpha (subjected to protein stability regulation) and constitutively expressed HIF-1beta. Besides hypoxia, diverse agonists are identified that stabilize HIF-1alpha during normoxia. Here we used a coculture system of RAW 264.7 macrophage cells and tubular LLC-PK(1) cells to establish that lipopolysaccharide- and interferon-gamma-stimulated but not resting macrophages elicited HIF-1alpha accumulation in LLC-PK(1) cells. Via pharmacological interventions such as blockade of nitric oxide (NO) production in macrophages, scavenging of NO with the use of 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, or application of tumor necrosis factor-alpha (TNF-alpha)-neutralizing antibodies, we identified NO and TNF-alpha as signaling molecules. Working in concert, NO and TNF-alpha have a stronger response when allowed direct cell-to-cell contact instead of contact with only the cell supernatant of activated macrophages. We show that signal transmission by NO with TNF-alpha in LLC-PK(1) cells is mediated via the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway, because it is blocked by wortmannin or dominant-negative forms of PI3-K as well as protein kinase B. We conclude that NO and TNF-alpha, derived from activated macrophages, provoke HIF-1alpha stabilization in LLC-PK(1) cells under normoxic conditions, which underscores HIF-1alpha stabilization due to intercellular regulation.
AB - Hypoxic/ischemic conditions provoke activation of the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 is composed of HIF-1alpha (subjected to protein stability regulation) and constitutively expressed HIF-1beta. Besides hypoxia, diverse agonists are identified that stabilize HIF-1alpha during normoxia. Here we used a coculture system of RAW 264.7 macrophage cells and tubular LLC-PK(1) cells to establish that lipopolysaccharide- and interferon-gamma-stimulated but not resting macrophages elicited HIF-1alpha accumulation in LLC-PK(1) cells. Via pharmacological interventions such as blockade of nitric oxide (NO) production in macrophages, scavenging of NO with the use of 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, or application of tumor necrosis factor-alpha (TNF-alpha)-neutralizing antibodies, we identified NO and TNF-alpha as signaling molecules. Working in concert, NO and TNF-alpha have a stronger response when allowed direct cell-to-cell contact instead of contact with only the cell supernatant of activated macrophages. We show that signal transmission by NO with TNF-alpha in LLC-PK(1) cells is mediated via the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway, because it is blocked by wortmannin or dominant-negative forms of PI3-K as well as protein kinase B. We conclude that NO and TNF-alpha, derived from activated macrophages, provoke HIF-1alpha stabilization in LLC-PK(1) cells under normoxic conditions, which underscores HIF-1alpha stabilization due to intercellular regulation.
KW - Animals
KW - Coculture Techniques
KW - Swine
KW - Transcription Factors/metabolism
KW - Inflammation Mediators/metabolism
KW - Signal Transduction/physiology
KW - Enzyme Inhibitors/pharmacology
KW - Tumor Necrosis Factor-alpha/metabolism
KW - Anoxia/metabolism/physiopathology
KW - Cell Communication/physiology
KW - Chemotaxis, Leukocyte/drug effects/physiology
KW - Epithelial Cells/drug effects/metabolism
KW - Free Radical Scavengers/pharmacology
KW - Hypoxia-Inducible Factor 1, alpha Subunit
KW - Inflammation/metabolism/physiopathology
KW - Kidney Tubules/drug effects/metabolism
KW - LLC-PK1 Cells
KW - Macrophages/drug effects/metabolism
KW - Nitric Oxide/metabolism
KW - Phosphatidylinositol 3-Kinases/antagonists & inhibitors/metabolism
KW - Protein-Serine-Threonine Kinases
KW - Proto-Oncogene Proteins/antagonists & inhibitors/metabolism
KW - Proto-Oncogene Proteins c-akt
KW - Animals
KW - Coculture Techniques
KW - Swine
KW - Transcription Factors/metabolism
KW - Inflammation Mediators/metabolism
KW - Signal Transduction/physiology
KW - Enzyme Inhibitors/pharmacology
KW - Tumor Necrosis Factor-alpha/metabolism
KW - Anoxia/metabolism/physiopathology
KW - Cell Communication/physiology
KW - Chemotaxis, Leukocyte/drug effects/physiology
KW - Epithelial Cells/drug effects/metabolism
KW - Free Radical Scavengers/pharmacology
KW - Hypoxia-Inducible Factor 1, alpha Subunit
KW - Inflammation/metabolism/physiopathology
KW - Kidney Tubules/drug effects/metabolism
KW - LLC-PK1 Cells
KW - Macrophages/drug effects/metabolism
KW - Nitric Oxide/metabolism
KW - Phosphatidylinositol 3-Kinases/antagonists & inhibitors/metabolism
KW - Protein-Serine-Threonine Kinases
KW - Proto-Oncogene Proteins/antagonists & inhibitors/metabolism
KW - Proto-Oncogene Proteins c-akt
M3 - SCORING: Journal article
VL - 284
SP - 439
EP - 446
JO - AM J PHYSIOL-CELL PH
JF - AM J PHYSIOL-CELL PH
SN - 0363-6143
IS - 2
M1 - 2
ER -
