Measurement of Soluble Epoxide Hydrolase (sEH) Activity

Morisseau, Christophe; Hammock, Bruce D.

doi:10.1002/0471140856.tx0423s33

Cited by 70 publications

(102 citation statements)

References 28 publications

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“…sEH is expressed in asthmatic lung (22) and plays important roles in arachidonic acid metabolism (24). Expression of sEH is increased by oxidative stress (21).…”

Section: Discussionmentioning

confidence: 99%

Lipoxin Generation Is Related to Soluble Epoxide Hydrolase Activity in Severe Asthma

Ono

Dutile

Kazani

et al. 2014

Am J Respir Crit Care Med

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Rationale: Severe asthma is characterized by airway inflammatory responses associated with aberrant metabolism of arachidonic acid. Lipoxins (LX) are arachidonate-derived pro-resolving mediators that are decreased in severe asthma, yet mechanisms for defective LX biosynthesis and a means to increase LXs in severe asthma remain to be established.Objectives: To determine if oxidative stress and soluble epoxide hydrolase (sEH) activity are linked to decreased LX biosynthesis in severe asthma.Methods: Aliquots of blood, sputum, and bronchoalveolar lavage fluid were obtained from asthma subjects for mediator determination. Select samples were exposed to t-butyl-hydroperoxide or sEH inhibitor (sEHI) before activation. Peripheral blood leukocyte-platelet aggregates were monitored by flow cytometry, and bronchial contraction was determined with cytokine-treated human lung sections.Measurements and Main Results: 8-Isoprostane levels in sputum supernatants were inversely related to LXA 4 in severe asthma (r = 20.55; P = 0.03) and t-butyl-hydroperoxide decreased LXA 4 and 15-epi-LXA 4 biosynthesis by peripheral blood leukocytes. LXA 4 and 15-epi-LXA 4 levels were inversely related to sEH activity in sputum supernatants and sEHIs significantly increased 14,15-epoxy-eicosatrienoic acid and 15-epi-LXA 4 generation by severe asthma whole blood and bronchoalveolar lavage fluid cells. The abundance of peripheral blood leukocyte-platelet aggregates was related to asthma severity. In a concentration-dependent manner, LXs significantly inhibited plateletactivating factor-induced increases in leukocyte-platelet aggregates (70.8% inhibition [LXA 4 100 nM], 78.3% inhibition [15-epi-LXA 4 100 nM]) and 15-epi-LXA 4 markedly inhibited tumor necrosis factora-induced increases in bronchial contraction.Conclusions: LX levels were decreased by oxidative stress and sEH activity. Inhibitors of sEH increased LXs that mediated antiphlogistic actions, suggesting a new therapeutic approach for severe asthma. Clinical trial registered with www.clinicaltrials.gov (NCT 00595114).

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“…sEH is expressed in asthmatic lung (22) and plays important roles in arachidonic acid metabolism (24). Expression of sEH is increased by oxidative stress (21).…”

Section: Discussionmentioning

confidence: 99%

Lipoxin Generation Is Related to Soluble Epoxide Hydrolase Activity in Severe Asthma

Ono

Dutile

Kazani

et al. 2014

Am J Respir Crit Care Med

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“…Sum EET+DHET and sum EET concentrations were calculated and normalized to cell density. In order to quantify cellular sEH metabolic activity, 25 ng 14,15-EET was incubated with 1 g total cell lysate from 3T3-L1 preadipocytes or mature adipocytes in a 0.1 ml volume of PBS plus 0.1 mg/ml BSA for 10 min at 37°C, as described ( 25 ). The reactions were stopped with 1 ml of ice-cold ethyl acetate, spiked with internal standard, and then evaporated to dryness under nitrogen gas.…”

Section: Cyp Epoxygenase and Seh Activity Assaymentioning

confidence: 99%

Functional characterization of cytochrome P450-derived epoxyeicosatrienoic acids in adipogenesis and obesity

Zha

Edin

Vendrov

et al. 2014

Journal of Lipid Research

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Obesity is a major public health problem that contributes to the development of type 2 diabetes and cardiovascular disease ( 1 ). Excessive lipid accumulation in adipose tissue is a key pathological driver of obesity, and is manifested by an increase in the number (hyperplasia) and size (hypertrophy) of adipocytes. Differentiation of preadipocytes to mature adipocytes (adipogenesis) is an integral mediator of this process, and is under the control of a transcription factor network that regulates lipid biosynthesis and metabolism ( 2 ). Most notably, PPAR-␥ , CCAAT/ enhancer-binding protein (C/EBP) ␣ , and sterol regulatoryelement-binding protein (SREBP)1c are central mediators of this process through the regulation of LPL, acetyl-CoA carboxylase 1 (ACC1), and FAS expression ( 2, 3 ). Sustained activation of this process, however, is a key pathological driver of obesity that results in adipocyte dysfunction, glucose intolerance, and ultimately the development of insulin resistance and type 2 diabetes ( 4, 5 ). Consequently, an improved understanding of the key pathways that regulate adipogenesis and adipocyte function offers enormous potential to facilitate the development of novel therapeutic strategies that mitigate the development and progression of obesity-associated metabolic diseases. Abstract

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“…In the end-point setup of the assay, one needed to proportionally increase the incubation time to still be able to signifi cantly measure activity above background as the enzyme is diluted. For routine assays, an sEH concentration of around 1-10 nM is normally used, and incubation for 5-10 min is usual ( 30 ). In this report, as low picomolar concentrations of enzymes were reached, incubation times of roughly 5,000 min ( ≈ 96 h) were needed to signifi cantly measure activity.…”

Section: Kinetic Constant Analysismentioning

confidence: 99%

Effect of soluble epoxide hydrolase polymorphism on substrate and inhibitor selectivity and dimer formation

Morisseau

Wecksler

Deng

et al. 2014

Journal of Lipid Research

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Measurement of Soluble Epoxide Hydrolase (sEH) Activity

Cited by 70 publications

References 28 publications

Lipoxin Generation Is Related to Soluble Epoxide Hydrolase Activity in Severe Asthma

Lipoxin Generation Is Related to Soluble Epoxide Hydrolase Activity in Severe Asthma

Functional characterization of cytochrome P450-derived epoxyeicosatrienoic acids in adipogenesis and obesity

Effect of soluble epoxide hydrolase polymorphism on substrate and inhibitor selectivity and dimer formation

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