Cytochrome P450 (CYP) 2J2 gene transfection attenuates MMP‐9 via inhibition of NF‐κβ 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.

Chronic Doxorubicin Cardiotoxicity Modulates Cardiac Cytochrome P450-Mediated Arachidonic Acid Metabolism in Rats

“…On the other hand, 20-HETE, an eicosanoid produced by P450 -hydroxylases, has been reported to have a detrimental effect on the heart (Chábová et al, 2007;Lv et al, 2008;Minuz et al, 2008), leading to cardiac hypertrophy and heart failure (Bao et al, 2011). Mechanistically, these opposing effects of P450-derived AA metabolites are mediated through several intracellular signaling cascades that have been implicated in development and/or progression of cardiovascular toxicity, e.g., nuclear factor B, mitogen-activated protein kinase, and matrix metalloproteinase-9 (Node et al, 1999;Sun et al, 1999;Elbekai and El-Kadi, 2006;Ishizuka et al, 2008;Moshal et al, 2008). As an important player affecting the balance between P450-derived AA metabolites, soluble epoxide hydrolase (sEH) enzyme hydrolyzes the cardioprotective metabolites EETs to biologically less active metabolites, dihydroxyeicosatrienoic acids (DHETs) (Imig et al, 2002).…”

Role of Copper and Homocysteine in Pressure Overload Heart Failure

“…Image analysis of the protein bands was performed using UMAX PowrLock II (Taiwan). The protein expression intensity was assessed by integrated optical density [14].…”

CYP2J2-Derived EETs Attenuated Angiotensin II-Induced Adventitial Remodeling via Reduced Inflammatory Response

“…EETs may be PPARγ ligand [16], and PPARγ agonists attenuated Ang II-induced inflammation via NF-κB pathway [48]. Additionally, EETs exerted anti-inflammatory effect via inhibiting IκBα degradation and NF-κB translocation [15]. Our experiments suggested a crucial role of PPARγ-NF-κB pathway in the anti-inflammatory effect of exogenous EETs administration and CYP2J2 overexpression.…”

“…Studies showed that PPARγ ligands suppress inflammation by inhibiting the activation of nuclear factor κB (NF-κB), activator protein 1 (AP-1) and signal transducers and activator of transcription (STAT). Moshal et al [15] reported that CYP2J2 transfection or exogenous 8,9-EET addition phosphorylated AKT and attenuated homocysteine-induced MMP-9 activation by inhibiting NF-κB nuclear translocation, NF-κB-DNA binding, and IκBα activation. Further studies indicated that PPARγ is an effector of EETs, as blocking PPARγ by GW9662 abolished EET/AUDA-mediated anti-inflammatory effect [16].…”