HNE interacts directly with JNK isoforms in human hepatic stellate cells.
4-Hydroxy-2,3-nonenal (HNE) is an aldehydic end product of lipid peroxidation which has been detected in vivo in clinical and experimental conditions of chronic liver damage. HNE has been shown to stimulate procollagen type I gene expression and synthesis in human hepatic stellate cells (hHSC) which are known to play a key role in liver fibrosis. In this study we investigated the molecular mechanisms underlying HNE actions in cultured hHSC. HNE, at doses compatible with those detected in vivo, lead to an early generation of nuclear HNE-protein adducts of 46, 54, and 66 kD, respectively, as revealed by using a monoclonal antibody specific for HNE-histidine adducts. This observation is related to the lack of crucial HNE-metabolizing enzymatic activities in hHSC. Kinetics of appearance of these nuclear adducts suggested translocation of cytosolic proteins. The p46 and p54 isoforms of c-Jun amino-terminal kinase (JNKs) were identified as HNE targets and were activated by this aldehyde. A biphasic increase in AP-1 DNA binding activity, associated with increased mRNA levels of c-jun, was also observed in response to HNE. HNE did not affect the Ras/ERK pathway, c-fos expression, DNA synthesis, or NF-kappaB binding. This study identifies a novel mechanism linking oxidative stress to nuclear signaling in hHSC. This mechanism is not based on redox sensors and is stimulated by concentrations of HNE compatible with those detected in vivo, and thus may be relevant during chronic liver diseases.
Parola
M
M
Dipartimento di Medicina ed Oncologia Sperimentale, University of Torino, Torino, Italy.
Robino
G
G
Marra
F
F
Pinzani
M
M
Bellomo
G
G
Leonarduzzi
G
G
Chiarugi
P
P
Camandola
S
S
Poli
G
G
Waeg
G
G
Gentilini
P
P
Dianzani
M U
MU
D000447:Aldehydes / Q000494:pharmacology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002478:Cells, Cultured; D016762:Genes, fos; D016758:Genes, jun; D006639:Histidine / Q000737:chemistry / Q000187:drug effects; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases; D015227:Lipid Peroxidation; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000209:etiology*; D008107:Liver Diseases / Q000150:complications / Q000378:metabolism*; D048670:MAP Kinase Kinase 4; D048052:Mitogen-Activated Protein Kinase 3; D020929:Mitogen-Activated Protein Kinase Kinases; D020928:Mitogen-Activated Protein Kinases; D008970:Molecular Weight; D018384:Oxidative Stress; D011494:Protein Kinases / Q000737:chemistry / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D018808:Transcription Factor AP-1 / Q000378:metabolism
D000447:Aldehydes; D018808:Transcription Factor AP-1; D006639:Histidine; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D020929:Mitogen-Activated Protein Kinase Kinases; C027576:4-hydroxy-2-nonenal