Determination of the Dominant Arachidonic Acid Cytochrome P450 Monooxygenases in Rat Heart, Lung, Kidney, and Liver: Protein Expression and Metabolite Kinetics

El-Sherbeni, Ahmed A.; Aboutabl, Mona E.; Zordoky, Beshay N.; Anwar-Mohamed, Anwar

doi:10.1208/s12248-012-9425-7

Cited by 39 publications

(31 citation statements)

References 46 publications

(55 reference statements)

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“…In line with the current findings, it has been reported that the expression of CYP2B1 and/or CYP2J3 was increased in SHR (Spontaneously hypertensive rat), pressure overload-induced cardiac hypertrophy, and isoproterenol-induced cardiac hypertrophy (Thum and Borlak, 2002;Althurwi et al, 2013;El-Sherbeni and El-Kadi, 2014). CYP2B1, CYP2C23, and CYP2J were all reported to produce EETs where CYP2C and CYP2J subfamilies appear to be the primary P450 epoxygenases, in rat and human heart, respectively (Roman, 2002;Imig, 2012;El-Sherbeni et al, 2013). EET levels are critically regulated by the sEH enzyme, as it catalyzes their degradation to DHETs, thus eliminating their biologic activity.…”

Section: P450s In the Development Of Cardiac Hypertrophysupporting

confidence: 89%

Early Changes in Cytochrome P450s and Their Associated Arachidonic Acid Metabolites Play a Crucial Role in the Initiation of Cardiac Hypertrophy Induced by Isoproterenol

et al. 2015

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Cytochrome P450 enzymes (P450s), along with their cardioprotective metabolites the epoxyeicosatrienoic acids (EETs) and cardiotoxic metabolite 20-hydroxyeicosatetraeonic acid (20-HETE), were found to be altered in cardiac hypertrophy; however, it is unclear whether these changes are causal or epiphenomenon. Therefore, we hypothesized that P450s and their metabolites play a crucial role in the initiation of cardiac hypertrophy. To test our hypothesis, rats and RL-14 cells were treated with the hypertrophic agonist isoproterenol for different time periods. Thereafter, in vivo heart function and wall thickness were assessed using echocardiography. Moreover, the role of P450 epoxygenases, v-hydroxylases, and soluble epoxide hydrolase (sEH) were determined at mRNA, protein, and activity levels using real-time polymerase chain reaction, Western blot, and liquid chromatography-mass spectrometry, respectively. Our results show that in vivo and in vitro hypertrophy was initiated after 72 hours and 6 hours of isoproterenol treatment, respectively. Studies performed at the prehypertrophy phase showed a significant decrease in P450 epoxygenases along with a significant induction of sEH activity. Consequently, lower EET and higher dihydroxyeicosatrienoic acid levels were observed during this phase. However, significant increases in P450 v-hydroxylase along with its associated metabolite, 20-HETE, were detected only in vivo. Interestingly, increasing EET levels by P450 epoxygenase induction, sEH inhibition, or exogenous administration of EET prevented the initiation of cardiac hypertrophy through a nuclear factor-kB-mediated mechanism. Taken together, these findings reveal a crucial role of P450 epoxygenases and EETs in the development of cardiac hypertrophy, which could uncover novel targets for prevention of heart failure at early stages.

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Section: P450s In the Development Of Cardiac Hypertrophysupporting

confidence: 89%

Early Changes in Cytochrome P450s and Their Associated Arachidonic Acid Metabolites Play a Crucial Role in the Initiation of Cardiac Hypertrophy Induced by Isoproterenol

et al. 2015

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“…This is in line with our results that the liver has the highest formation rate of CYP1As major metabolites, v-1→4 HETEs, and the heart has the lowest rate. The expression of CYP2Cs has been reported to be the highest in the liver, followed by the kidneys, and it is lowest in the heart and lungs (El-Sherbeni et al, 2013). This finding correlates with the rate of EETs formation, which is highest in the liver, followed by kidneys, then the heart and lungs.…”

Section: Discussionmentioning

confidence: 94%

“…P450 expression and their catalytic activities are the two factors that dictate the contribution of a P450 enzyme to AA metabolism in a tissue. With respect to P450 dmd.aspetjournals.org expression, substantial differences have been reported between different tissues and organs (Zordoky et al, 2008;El-Sherbeni et al, 2013). Additionally, AA-metabolizing activity demonstrated great variation among P450 enzymes.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Arachidonic Acid Metabolism by Rat Cytochrome P450 Enzymes: The Involvement of CYP1As

El-Sherbeni

2014

Drug Metab Dispos

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Cytochrome P450 (P450) enzymes mediate arachidonic acid (AA) oxidation to several biologically active metabolites. Our aims in this study were to characterize AA metabolism by different recombinant rat P450 enzymes and to identify new targets for modulating P450-AA metabolism in vivo. A liquid chromatography-mass spectrometry method was developed and validated for the simultaneous measurements of AA and 15 of its P450 metabolites. CYP1A1, CYP1A2, CYP2B1, CYP2C6, and CYP2C11 were found to metabolize AA with high catalytic activity, and CYP2A1, CYP2C13, CYP2D1, CYP2E1, and CYP3A1 had lower activity. CYP1A1 and CYP1A2 produced v-1→4 hydroxyeicosatetraenoic acids (HETEs) as 88.7 and 62.7%, respectively, of the total metabolites formed. CYP2C11 produced epoxyeicosatrienoic acids (EETs) as 61.3%, and CYP2C6 produced midchain HETEs and EETs as 48.3 and 29.4%, respectively, of the total metabolites formed. The formation of CYP1A1, CYP1A2, CYP2C6, and CYP2C11 major metabolites followed an atypical kinetic profile of substrate inhibition. CYP1As inhibition by a-naphthoflavone or anti-CYP1As antibodies significantly reduced v-1→4 HETE formation in the lungs and liver, whereas CYP1As induction by 3-methylcholanthrene resulted in a significant increase in v-1→4 HETEs formation in the heart, lungs, kidney, and livers by 370, 646, 532, and 848%, respectively. In conclusion, our results suggest that CYP1As and CYP2Cs are major players in the metabolism of AA. The significant contribution of CYP1As to AA metabolism and their strong inducibility suggest their possible use as targets for the prevention and treatment of several diseases.

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“…The results of this study found that 14,15-EEZE dose-dependently reversed the CP's To investigate why AA P450 enzyme metabolism was influenced by multiple doses of the CP, the expressions and catalyzing activities of AA-related P450 enzymes in the rat heart were studied. CYP2C11 and CYP2J3 are two main AA epoxygenases in the rat heart (El-Sherbeni et al, 2013). Cardiac CYP1A1, CYP2B1, and CYP2E1 have also been reported to display AA epoxygenase activity (El-Sherbeni and El-Kadi, 2014).…”

Section: Discussionmentioning

confidence: 99%

Cardiotonic Pill Reduces Myocardial Ischemia-Reperfusion Injury via Increasing EET Concentrations in Rats

Hao

Jiang

et al. 2016

Drug Metabolism and Disposition

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Accumulating data suggest that epoxyeicosatrienoic acids (EETs) and 20-hydroxyeicosatetraenoic acid, both cytochrome P450 (P450) enzyme metabolites of arachidonic acid (AA), play important roles in cardiovascular diseases. For many years, the cardiotonic pill (CP), an herbal preparation derived from Salviae Miltiorrhizae Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Borneolum Syntheticum, has been widely used in China for the treatment of coronary artery disease. However, its pharmacological mechanism has not been well elucidated. The purpose of this study was to investigate the chronic effects of the CP on myocardial ischemia-reperfusion injury (MIRI) and AA P450 enzyme metabolism in rats (in vivo) and H9c2 cells (in vitro). The results showed that CP dose dependently (10, 20, and 40 mg/kg/d; 7 days) mitigated MIRI in rats. The plasma concentrations of EETs in CP-treated ischemia-reperfusion (I/R) rats (40 mg/kg/d; 7 days) were significantly higher (P < 0.05) than those in controls. Cardiac Cyp1b1, Cyp2b1, Cyp2e1, Cyp2j3, and Cyp4f6 were significantly induced (P < 0.05); CYP2J and CYP2C11 proteins were upregulated (P < 0.05); and AA-epoxygenases activity was significantly increased (P < 0.05) after CP (40 mg/kg/d; 7 days) administration in rats. In H9c2 cells, the CP also increased (P < 0.05) the EET concentrations and showed protection in hypoxia-reoxygenation (H/R) cells. However, an antagonist of EETs, 14,15-epoxyeicosa-5(Z)-enoic acid, displayed a dose-dependent depression of the CP's protective effects in H/R cells. In conclusion, upregulation of cardiac epoxygenases after multiple doses of the CP-leading to elevated concentrations of cardioprotective EETs after myocardial I/R-may be the underlying mechanism, at least in part, for the CP's cardioprotective effect in rats.

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Determination of the Dominant Arachidonic Acid Cytochrome P450 Monooxygenases in Rat Heart, Lung, Kidney, and Liver: Protein Expression and Metabolite Kinetics

Cited by 39 publications

References 46 publications

Early Changes in Cytochrome P450s and Their Associated Arachidonic Acid Metabolites Play a Crucial Role in the Initiation of Cardiac Hypertrophy Induced by Isoproterenol

Early Changes in Cytochrome P450s and Their Associated Arachidonic Acid Metabolites Play a Crucial Role in the Initiation of Cardiac Hypertrophy Induced by Isoproterenol

Characterization of Arachidonic Acid Metabolism by Rat Cytochrome P450 Enzymes: The Involvement of CYP1As

Cardiotonic Pill Reduces Myocardial Ischemia-Reperfusion Injury via Increasing EET Concentrations in Rats

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