Cytochrome P450 (CYP) 2J2 gene transfection attenuates MMP-9 via inhibition of NF-kappabeta in hyperhomocysteinemia.

Abstract

Hyperhomocysteinemia (HHcy) is associated with atherosclerotic events involving the modulation of arachidonic acid (AA) metabolism and the activation of matrix metalloproteinase-9 (MMP-9). Cytochrome P450 (CYP) epoxygenase-2J2 (CYP2J2) is abundant in the heart endothelium, and its AA metabolites epoxyeicosatrienoic acids (EETs) mitigates inflammation through NF-kappabeta. However, the underlying molecular mechanisms for MMP-9 regulation by CYP2J2 in HHcy remain obscure. We sought to determine the molecular mechanisms by which P450 epoxygenase gene transfection or EETs supplementation attenuate homocysteine (Hcy)-induced MMP-9 activation. CYP2J2 was over-expressed in mouse aortic endothelial cells (MAECs) by transfection with the pcDNA3.1/CYP2J2 vector. The effects of P450 epoxygenase transfection or exogenous supplementation of EETs on NF-kappabeta-mediated MMP-9 regulation were evaluated using Western blot, in-gel gelatin zymography, electromobility shift assay, immunocytochemistry. The result suggested that Hcy downregulated CYP2J2 protein expression and dephosphorylated PI3K-dependent AKT signal. Hcy induced the nuclear translocation of NF-kappabeta via downregulation of IKbetaalpha (endogenous cytoplasmic inhibitor of NF-kappabeta). Hcy induced MMP-9 activation by increasing NF-kappabeta-DNA binding. Moreover, P450 epoxygenase transfection or exogenous addition of 8,9-EET phosphorylated the AKT and attenuated Hcy-induced MMP-9 activation. This occurred, in part, by the inhibition of NF-kappabeta nuclear translocation, NF-kappabeta-DNA binding and activation of IKbetaalpha. The study unequivocally suggested the pivotal role of EETs in the modulation of Hcy/MMP-9 signal.

• Authors: Karni S Moshal, Darryl C Zeldin, Srinivas D Sithu, Utpal Sen, Neetu Tyagi, Munish Kumar, William M Hughes Jr, Naira Metreveli, Dorothea S E Rosenberger, Mahavir Singh, Thomas P Vacek, Walter E Rodriguez, Adeagbo Ayotunde, Suresh C Tyagi
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 215 Issue 3 Pg. 771-81 (Jun 2008) ISSN: 1097-4652 [Electronic] United States
• PMID: 18181170 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)
• Copyright: (c) 2008 Wiley-Liss, Inc.

Chemical References

• I-kappa B Proteins
• Nfkbia protein, mouse
• Transcription Factor RelA
• Homocysteine
• NF-KappaB Inhibitor alpha
• 8,9-epoxyeicosatrienoic acid
• Cytochrome P-450 Enzyme System
• Oxygenases
• Cytochrome P-450 CYP2J2
• Proto-Oncogene Proteins c-akt
• Matrix Metalloproteinase 9
• 8,11,14-Eicosatrienoic Acid

Topics

• 8,11,14-Eicosatrienoic Acid (analogs & derivatives, pharmacology)
• Animals
• Cells, Cultured
• Cytochrome P-450 CYP2J2
• Cytochrome P-450 Enzyme System (genetics, metabolism)
• Enzyme Activation (drug effects)
• Enzyme Induction (drug effects)
• Homocysteine (pharmacology)
• Hyperhomocysteinemia (enzymology, metabolism)
• I-kappa B Proteins (metabolism)
• Matrix Metalloproteinase 9 (biosynthesis, metabolism)
• Mice
• NF-KappaB Inhibitor alpha
• Oxygenases (genetics, metabolism)
• Phosphatidylinositol 3-Kinases (metabolism)
• Phosphorylation (drug effects)
• Proto-Oncogene Proteins c-akt (metabolism)
• Transcription Factor RelA (antagonists & inhibitors, metabolism)
• Transfection