title: The effect of peroxynitrite decomposition catalyst MnTBAP on aldehyde dehydrogenase-2 nitration by organic nitrates: role in nitrate tolerance.
abstract: Bioconversion of glyceryl trinitrate (GTN) into nitric oxide (NO) by aldehyde dehydrogenase-2 (ALDH-2) is a crucial mechanism which drives vasodilatory and antiplatelet effect of organic nitrates in vitro and in vivo. Oxidative stress generated by overproduction of free radical species, mostly superoxide anions and NO-derived peroxynitrite, has been suggested to play a pivotal role in the development of nitrate tolerance, though the mechanism still remains unclear. Here we studied the free radical-dependent impairment of ALDH-2 in platelets as well as vascular tissues undergoing organic nitrate ester tolerance and potential benefit when using the selective peroxynitrite decomposition catalyst Mn(III) tetrakis (4-Benzoic acid) porphyrin (MnTBAP). Washed human platelets were made tolerant to nitrates via incubation with GTN for 4h. This was expressed by attenuation of platelet aggregation induced by thrombin (40U/mL), an effect accompanied by GTN-related induction of cGMP levels in platelets undergoing thrombin-induced aggregation. Both effects were associated to attenuated GTN-induced nitrite formation in platelets supernatants and to prominent nitration of ALDH-2, the GTN to NO metabolizing enzyme, suggesting that GTN tolerance was associated to reduced NO formation via impairment of ALDH-2. These effects were all antagonized by co-incubation of platelets with MnTBAP, which restored GTN-induced responses in tolerant platelets. Comparable effect was found under in in vivo settings. Indeed, MnTBAP (10mg/kg, i.p.) significantly restored the hypotensive effect of bolus injection of GTN in rats made tolerants to organic nitrates via chronic administration of isosorbide-5-mononitrate (IS-5-MN), thus confirming the role of peroxynitrite overproduction in the development of tolerance to vascular responses induced by organic nitrates. In conclusion, oxidative stress subsequent to prolonged use of organic nitrates, which occurs via nitration of ALDH-2, represents a key event in GTN tolerance, an effect counteracted both in vitro and in vivo by novel peroxynitrite decomposition catalyst.
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1); IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166 Rome, Italy(2). Electronic address: mollace@libero.it.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Mollace
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1); IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166 Rome, Italy(2). Electronic address: muscoli@unicz.it.
authors.forename: Carolina
authors.initials: C
authors.lastname: Muscoli
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: c_dagostino@libero.it.
authors.forename: Concetta
authors.initials: C
authors.lastname: Dagostino
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: luigi.giancotti@unicz.it.
authors.forename: Luigino Antonio
authors.initials: LA
authors.lastname: Giancotti
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1); IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166 Rome, Italy(2). Electronic address: micaela.gliozzi@gmail.com.
authors.forename: Micaela
authors.initials: M
authors.lastname: Gliozzi
authors.affiliation: ARPACAL, Viale Lungomare Loc. Mosca snc, 88063 Catanzaro Lido, Italy(3). Electronic address: i.sacco@arpacal.it.
authors.forename: Iolanda
authors.initials: I
authors.lastname: Sacco
authors.affiliation: ARPACAL, Viale Lungomare Loc. Mosca snc, 88063 Catanzaro Lido, Italy(3). Electronic address: vvisalli@libero.it.
authors.forename: Valeria
authors.initials: V
authors.lastname: Visalli
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: gratteri@unicz.it.
authors.forename: Santo
authors.initials: S
authors.lastname: Gratteri
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: palma@unicz.it.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Palma
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: nataliamalara@unicz.it.
authors.forename: Natalia
authors.initials: N
authors.lastname: Malara
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: xabaras3@hotmail.com.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Musolino
authors.affiliation: Interregional Research Center for Food Safety & Health (IRC-FSH), Department of Health Science, University "Magna Graecia" of Catanzaro, Complesso "Ninì Barbieri", 88021 Roccelletta di Borgia, Italy(1). Electronic address: carresi@unicz.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Carresi
authors.affiliation: Department of Internal Medicine, Division of Cardiology, University Hospital of Tor Vergata, Viale Oxford, 81, 00133 Rome, Italy(4). Electronic address: saveriomuscoli@gmail.com.
authors.forename: Saverio
authors.initials: S
authors.lastname: Muscoli
authors.affiliation: IRCCS San Raffaele Pisana, Via di Val Cannuta, 247, 00166 Rome, Italy(2). Electronic address: cristiana.vitale@sanraffaele.it.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Vitale
authors.affiliation: Department of Pharmacological and Physiological Science, Saint. Louis University School of Medicine, 1402 South Grand Blvd, 63104 MO, USA(5). Electronic address: salvemd@slu.edu.
authors.forename: Daniela
authors.initials: D
authors.lastname: Salvemini
authors.affiliation: Department of Internal Medicine, Division of Cardiology, University Hospital of Tor Vergata, Viale Oxford, 81, 00133 Rome, Italy(4). Electronic address: romeocerabino@tin.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Romeo
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D004361:Drug Tolerance; D004730:Endothelium, Vascular / Q000187:drug effects; D006801:Humans; D007022:Hypotension / Q000139:chemically induced; D007548:Isosorbide Dinitrate / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D008665:Metalloporphyrins / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D005996:Nitroglycerin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects; D051381:Rats; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000096:biosynthesis; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D008665:Metalloporphyrins; D009566:Nitrates; D009573:Nitrites; D014665:Vasodilator Agents; C097284:manganese(III)-tetrakis(4-benzoic acid)porphyrin; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; D000444:Aldehyde Dehydrogenase; D013917:Thrombin; D005996:Nitroglycerin; D006152:Cyclic GMP; D007548:Isosorbide Dinitrate; C030397:isosorbide-5-mononitrate
keywords: ALDH-2 nitration; Glyceryl trinitrate (GTN) (PubChem CID: 4510); Isosorbide-5-mononitrate (IS-5-MN) (PubChem CID: 27661); Mn(III)tetrakis(4-Benzoic acid) porphyrin (MnTBAP) (PubChem CID: 16760566); Nitrate tolerance; Nitric oxide (NO) (PubChem CID: 145068); Oxidative stress; Peroxynitrite decomposition catalyst