On (N.A.M.)Received June 20, 2013; accepted September 10,ABSTRACT Cytochrome P450 2J2 plays a IL-15 Protein Purity & Documentation considerable part within the epoxidation of arachidonic acid to signaling molecules crucial in cardiovascular events. CYP2J2 also contributes to drug metabolism and is accountable for the intestinal clearance of ebastine. However, the interaction involving arachidonic acid metabolism and drug metabolism in cardiac tissue, the primary expression website of CYP2J2, has not been examined. Here we investigate an adult-derived human main cardiac cell line as a suitable model to study metabolic drug interactions (inhibition and induction) of CYP2J2 in cardiac tissue. The primary human cardiomyocyte cell line demonstrated comparable mRNA-expression profiles of P450 enzymes to adult human ventricular tissue. CYP2J2 was the dominant IL-34 Protein manufacturer isozyme with minor contributions from CYP2D6 and CYP2E1. Both terfenadine and astemizole oxidation have been observed in this cell line, whereas midazolam was not metabolized suggesting lack of CYP3A activity. Compared with recombinant CYP2J2, terfenadine was hydroxylated in cardiomyocytes at a similar Km worth of 1.5 mM. The Vmax of terfenadine hydroxylation in recombinant enzyme was discovered to be 29.four pmol/pmol P450 per minute and in the cells six.0 pmol/pmol P450 per minute. CYP2J2 activity inside the cell line was inhibited by danazol, astemizole, and ketoconazole in submicromolar range, but also by xenobiotics recognized to result in cardiac adverse effects. From the 14 compounds tested for CYP2J2 induction, only rosiglitazone improved mRNA expression, by 1.8-fold. This cell model is often a helpful in vitro model to investigate the function of CYP2J2-mediated drug metabolism, arachidonic acid metabolism, and their association to drug induced cardiotoxicity.Introduction Cytochrome P450 2J2 has attracted particular attention for its capability to epoxidize arachidonic acid regioselectively to five,6-, 8,9-, 11,12-, or 14,15-epoxyeicosatrienoic acids (EETs) (Roman, 2002). These EETs have a lot of biological functions including, but not restricted to, angiogenesis, regulation of vasodilation, inhibition of cytokine-induced endothelial cell adhesion-molecule expression, inhibition of vascular smooth muscle cell migration, protection of endothelial cells against hypoxia-reoxygenation injury, upregulation of endothelial nitric oxide biosynthesis, and protection of doxorubicin-induced cardiotoxicity (Larsen et al., 2007; Spector and Norris, 2007; Yang et al., 2009; Zhang et al., 2009; Campbell and Fleming, 2010; Pfister et al., 2010). All these events are involved in cardiac electrophysiology and protect the heart from ischemic-reperfusion injury (Spiecker and Liao, 2006). A lot more particularly, the regioisomer 11,12-EET has been shown to be a potent activator in the ion channels sensitive to ATP, to directly reduce the membrane action possible in rat myocytes (Lu et al., 2001), and to boost recovery of ventricular repolarization following ischemia reperfusion injury (Batchu et al., 2009). These investigations drastically enhanced interest in CYP2J2 with regard to its enzymology, localized expression, plus the have to have for an in vitro model system suitable for studying the enzyme’s significance in keeping cardiomyocyte homeostasis.This operate was supported by the National Institutes of Health National Heart, Lung and Blood Institute [R01HL096706]. dx.doi.org/10.1124/dmd.113.053389. s This short article has supplemental material accessible at dmd.aspetjournals.org.CYP2J2 is predominantly expres.
