Development of a semi-automated LC/MS/MS method for the simultaneous quantitation of 14,15-epoxyeicosatrienoic acid, 14,15-dihydroxyeicosatrienoic acid, leukotoxin and leukotoxin diol in human plasma as biomarkers of soluble epoxide hydrolase activity in vivo

We tested the hypothesis that suppression of epoxyeicosatrienoic acid (EET) metabolism via genetic knockout of the gene for soluble epoxide hydrolase (sEH-KO), or female-specific downregulation of sEH expression, plays a role in the potentiation of pulmonary hypertension. We used male (M) and female (F) wild-type (WT) and sEH-KO mice; the latter have high pulmonary EETs. Right ventricular systolic pressure (RVSP) and mean arterial blood pressure (MABP) in control and in response to in vivo administration of U46619 (thromboxane analog), 14,15-EET, and 14,15-EEZE [14,15-epoxyeicosa-5(z)-enoic acid; antagonist of EETs] were recorded. Basal RVSP was comparable among all groups of mice, whereas MABP was significantly lower in F-WT than M-WT mice and further reduced predominantly in F-KO compared with M-KO mice. U46619 dose dependently increased RVSP and MABP in all groups of mice. The increase in RVSP was significantly greater and coincided with smaller increases in MABP in M-KO and F-WT mice compared with M-WT mice. In F-KO mice, the elevation of RVSP by U46619 was even higher than in M-KO and F-WT mice, associated with the least increase in MABP. 14,15-EEZE prevented the augmentation of U46619-induced elevation of RVSP in sEH-KO mice, whereas 14,15-EET-induced pulmonary vasoconstriction was comparable in all groups of mice. sEH expression in the lungs was reduced, paralleled with higher levels of EETs in F-WT compared with M-WT mice. In summary, EETs initiate pulmonary vasoconstriction but act as vasodilators systemically. High pulmonary EETs, as a function of downregulation or deletion of sEH, potentiate U46619-induced increases in RVSP in a female-susceptible manner.

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Assessment of soluble epoxide hydrolase activity in vivo: A metabolomic approach

Prostaglandins & Other Lipid Mediators ◽

10.1016/j.prostaglandins.2020.106410 ◽

2020 ◽

Vol 148 ◽

pp. 106410

Author(s):

Darko Stefanovski ◽

Pei-an Betty Shih ◽

Bruce D. Hammock ◽

Richard M. Watanabe ◽

Jang H. Youn

Keyword(s):

Epoxide Hydrolase ◽

Soluble Epoxide Hydrolase ◽

Hydrolase Activity ◽

Metabolomic Approach

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Elevated 14,15- epoxyeicosatrienoic acid by increasing of cytochrome P450 2C8, 2C9 and 2J2 and decreasing of soluble epoxide hydrolase associated with aggressiveness of human breast cancer

BMC Cancer ◽

10.1186/1471-2407-14-841 ◽

2014 ◽

Vol 14 (1) ◽

Cited By ~ 33

Author(s):

Xiaolong Wei ◽

Donghong Zhang ◽

Xiaowei Dou ◽

Na Niu ◽

Wenhe Huang ◽

...

Keyword(s):

Breast Cancer ◽

Cytochrome P450 ◽

Human Breast Cancer ◽

Epoxide Hydrolase ◽

Soluble Epoxide Hydrolase ◽

Epoxyeicosatrienoic Acid ◽

Human Breast ◽

Cytochrome P450 2C8

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Inhibitory Activity of Quercetin 3-O-Arabinofuranoside and 2-Oxopomolic Acid Derived from Malus domestica on Soluble Epoxide Hydrolase

Molecules ◽

10.3390/molecules25184352 ◽

2020 ◽

Vol 25 (18) ◽

pp. 4352

Author(s):

In Sook Cho ◽

Jang Hoon Kim ◽

Yunjia Lin ◽

Xiang Dong Su ◽

Jong Seong Kang ◽

...

Keyword(s):

Malus Domestica ◽

Epoxide Hydrolase ◽

Soluble Epoxide Hydrolase ◽

Corosolic Acid ◽

Allosteric Sites ◽

Ic50 Values ◽

Cardiovascular Tests ◽

Potential Inhibitors ◽

Molecular Docking And Dynamics

Flavonoids and triterpenoids were revealed to be the potential inhibitors on soluble epoxide hydrolase (sEH). The aim of this study is to reveal sEH inhibitors from Fuji apples. A flavonoid and three triterpenoids derived from the fruit of Malus domestica were identified as quercetin-3-O-arabinoside (1), ursolic acid (2), corosolic acid (3), and 2-oxopomolic acid (4). They had half-maximal inhibitory concentration of the inhibitors (IC50) values of 39.3 ± 3.4, 84.5 ± 9.5, 51.3 ± 4.9, and 11.4 ± 2.7 μM, respectively, on sEH. The inhibitors bound to allosteric sites of enzymes in mixed (1) and noncompetitive modes (2–4). Molecular simulations were carried out for inhibitors 1 and 4 to calculate the binding force of ligands to receptors. The inhibitors bound to the left (1) and right (4) pockets next to the enzyme’s active site. Based on analyses of their molecular docking and dynamics, it was shown that inhibitors 1 and 4 can stably bind sEH at 1 bar and 300 K. Finally, inhibitors 1 and 4 are promising candidates for further studies using cell-based assays and in vivo cardiovascular tests.

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In vitro and in vivo Metabolism of a Potent Inhibitor of Soluble Epoxide Hydrolase, 1-(1-Propionylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea

Frontiers in Pharmacology ◽

10.3389/fphar.2019.00464 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 5

Author(s):

Debin Wan ◽

Jun Yang ◽

Cindy B. McReynolds ◽

Bogdan Barnych ◽

Karen M. Wagner ◽

...

Keyword(s):

Epoxide Hydrolase ◽

Potent Inhibitor ◽

Soluble Epoxide Hydrolase ◽

In Vivo Metabolism

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Inhibition of soluble epoxide hydrolase preserves cardiomyocytes: role of STAT3 signaling

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00533.2009 ◽

2010 ◽

Vol 298 (2) ◽

pp. H679-H687 ◽

Cited By ~ 26

Author(s):

Matthias J. Merkel ◽

Lijuan Liu ◽

Zhiping Cao ◽

William Packwood ◽

Jennifer Young ◽

...

Keyword(s):

Epoxide Hydrolase ◽

Glucose Deprivation ◽

Soluble Epoxide Hydrolase ◽

Nucleotide Polymorphisms ◽

Gene Encoding ◽

Mutant Proteins ◽

Stat3 Signaling ◽

Stat3 Inhibition

Soluble epoxide hydrolase (sEH) metabolizes epoxyeicosatrienoic acids (EETs), primarily 14,15-EET. EETs are derived from arachidonic acid via P-450 epoxygenases and are cardioprotective. We tested the hypothesis that sEH deficiency and pharmacological inhibition elicit tolerance to ischemia via EET-mediated STAT3 signaling in vitro and in vivo. In addition, the relevance of single nucleotide polymorphisms (SNPs) of EPHX2 (the gene encoding sEH) on tolerance to oxygen and glucose deprivation and reoxygenation and glucose repletion (OGD/RGR) was assessed in male C57BL\6J (WT) or sEH knockout (sEHKO) cardiomyocytes by using transactivator of transcription (TAT)-mediated transduction with sEH mutant proteins. Cell death and hydrolase activity was lower in Arg287Gln EPHX2 mutants vs. nontransduced controls. Excess 14,15-EET and SEH inhibition did not improve cell survival in Arg287Gln mutants. In WT cells, the putative EET receptor antagonist, 14,15-EEZE, abolished the effect of 14,15-EET and sEH inhibition. Cotreatment with 14,15-EET and SEH inhibition did not provide increased protection. In vitro, STAT3 inhibition blocked 14,15-EET cytoprotection, but not the effect of SEH inhibition. However, STAT3 small interfering RNA (siRNA) abolished cytoprotection by 14,15-EET and sEH inhibition, but cells pretreated with JAK2 siRNA remained protected. In vivo, STAT3 inhibition abolished 14,15-EET-mediated infarct size reduction. In summary, the Arg287Gln mutation is associated with improved tolerance against ischemia in vitro, and inhibition of sEH preserves cardiomyocyte viability following OGD/RGR via an EET-dependent mechanism. In vivo and in vitro, 14,15-EET-mediated protection is mediated in part by STAT3.

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