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Homocy.teine and copper interact to promote ty.e 5 phosphodiesterase expression in rabbit cavernosal smooth muscle cells

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Asian Journal of Andrology 2008年第6期10卷 905-913页

作者： Matthew Hotston jamie y. jeremy Jonathon Bloor Nick S. Greaves Raj Persad Gianni Angelini Nilima Shukla Department of Urology.University.of BristoL Bristol BS8 1TH UK Department of Cardiac Surgery University of BristoL Bristol BS8 1TH UK

Aim： To study.the effects of homocy.teine and copper on ty.e 5 phosphodiesterase （PDE5） expression in cavemosai vascular smooth muscle cells （CVSMCs） and to investigate superoxide （O2-） derived from nicotinamide adenine dinucleotide phosphate oxidase as homocy.teine and copper generate O2-, and O2- upregulates PDE5 expression. Methods： CVSMCs derived from rabbit penis were incubated with homocy.teine or copper chloride with or without superoxide dismutase （SOD）, catalase, sildenafil citrate, or apocy.in （nicotinamide adenine dinucleotide phosphate inhibitor） for 16 h. The expression of PDE5 and of gly.eraldehy.e-3-phosphate dehy.rogenase （internal standard） was assessed using Western blot analy.is. In parallel, O2-was measured spectrophotometrically. Results： CuCl2 alone （up to 10 μmol/L） and homocy.teine alone （up to 100 μmol/L） had no effect on O2- formation in CVSMCs compared to controls. In combination, however, homocy.teine and CuCl2 markedly.increased O2- formation, an effect blocked by.SOD, catalase, apocy.in, and sildenafil （1 μmol/L） when co-incubated over the same time course. PDE5 expression was also significantly.increased in CVSMCs incubated with homocy.teine and CuCl2, compared to controls. This effect was also negated by.16-h co-incubation with SOD, catalase, apocy.in and sildenafil. Conclusion： This represents a novel pathogenic mechanism underly.ng ED, and indicates that the therapeutic actions of prolonged sildenafil use are mediated in part through inhibition of this pathway.

关键词： erectile dy.function superoxide PDE5 sildenafil

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Kinetic Characterization of Na,K-ATPase Inhibition by.the Acetaminophen Metabolite N-Acety.benzoquinoneimine

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Open Journal of Molecular and Integrative Phy.iology 2015年第1期5卷 1-17页

作者： Jeff B. Helms Lauren P. Saunders jamie Mey.r Charles J. Costa Eric Plowman Nick Williford Matthew Corbitt jeremy P. Holden Craig Gatto Department of Biology Upper Iowa University Fayette IA USA Division of Neuroscience and Phy.iology Department of Biological Science Illinois State University Normal IL USA Department of Internal Medicine University of Wisconsin-Madison Medical Center Madison WI USA

N-acety.benzoquinoneimine (NABQI) is a toxic metabolite of the common analgesic acetaminophen (APAP). NABQI is an electrophilic intermediate formed via the oxidation of APAP within the cy.ochrome P450 sy.tem. Within the normally.recommended low-dose use of APAP, NABQI is a minor metabolite which is either quickly.reduced back to APAP or conjugated to Glutathione (GSH) producing an innocuous by.product. However, with overdose or prolonged high-dose usage of acetaminophen, GSH levels can become depleted and the bioactive NABQI is thought to form adducts with proteins and oxidize protein sulfhy.ry.s producing intra- and intermolecular disulfide bridges in proteins. In this work we investigated the effect of NABQI on purified kidney.Na,K-ATPase to see if the clinical renal insufficiencies seen in APAP overdose may.be linked to inhibition of the Na,K-ATPase. Our work has shown that NABQI does indeed inhibit the Na,K-ATPase in a dose dependent (IC50 = 19.8 ± 2.9 μM) and irreversible manner. Interestingly. brief storage of NABQI at -20°C eliminates the irreversible effects of the compound, and leads to a product that remains a potent reversible inhibitor of the Na,K-ATPase (IC50 = 58.7 ± 19.5 μM). Further, the reversible inhibition produced by.stored NABQI competes with para-nitrop.

关键词： Na K-ATPase Paracetamol N-Acety.benzoquinoneimine Overdose Renal Electrophillic

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