title: Contribution of RhoA/Rho-kinase/MEK1/ERK1/2/iNOS pathway to ischemia/reperfusion-induced oxidative/nitrosative stress and inflammation leading to distant and target organ injury in rats.
abstract: The small G protein RhoA and its downstream effector Rho-kinase play an important role in various physiopathological processes including ischemia/reperfusion (I/R) injury. Reactive oxygen and nitrogen species produced by iNOS and NADPH oxidase are important mediators of inflammation and organ injury following an initial localized I/R event. The aim of this study was to determine whether RhoA/Rho-kinase signaling pathway increases the expression and activity of MEK1, ERK1/2, iNOS, gp91(phox), and p47(phox), and peroxynitrite formation which result in oxidative/nitrosative stress and inflammation leading to hindlimb I/R-induced injury in kidney as a distant organ and gastrocnemius muscle as a target organ. I/R-induced distant and target organ injury was performed by using the rat hindlimb tourniquet model. I/R caused an increase in the expression and/or activity of RhoA, MEK1, ERK1/2, iNOS, gp91(phox), p47(phox), and 3-nitrotyrosine and nitrotyrosine levels in the tissues. Although Rho-kinase activity was increased by I/R in the kidney, its activity was decreased in the muscle. Serum and tissue MDA levels and MPO activity were increased following I/R. I/R also caused an increase in SOD and catalase activities associated with decreased GSH levels in the tissues. Y-27632, a selective Rho-kinase inhibitor, (100µg/kg, i.p.; 1h before reperfusion) prevented the I/R-induced changes except Rho-kinase activity in the muscle. These results suggest that activation of RhoA/Rho-kinase/MEK1/ERK1/2/iNOS pathway associated with oxidative/nitrosative stress and inflammation contributes to hindlimb I/R-induced distant organ injury in rats. It also seems that hindlimb I/R induces target organ injury via upregulation of RhoA/MEK1/ERK1/2/iNOS pathway associated with decreased Rho-kinase activity.
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: A Nihal
authors.initials: AN
authors.lastname: Sari
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: Meltem
authors.initials: M
authors.lastname: Kacan
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: Demet
authors.initials: D
authors.lastname: Unsal
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: Seyhan
authors.initials: S
authors.lastname: Sahan Firat
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: C
authors.initials: C
authors.lastname: Kemal Buharalioglu
authors.affiliation: Department of Cardiovascular Surgery, Faculty of Medicine, Mersin University, Mersin, Turkey.
authors.forename: Ozden
authors.initials: O
authors.lastname: Vezir
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: Belma
authors.initials: B
authors.lastname: Korkmaz
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey.
authors.forename: Tuba
authors.initials: T
authors.lastname: Cuez
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mersin University, Mersin, Turkey.
authors.forename: Necmiye
authors.initials: N
authors.lastname: Canacankatan
authors.affiliation: Department of Cardiovascular Surgery, Faculty of Medicine, Mersin University, Mersin, Turkey.
authors.forename: Nehir
authors.initials: N
authors.lastname: Sucu
authors.affiliation: Department of Medicinal Biochemistry, Faculty of Medicine, Mersin University, Mersin, Turkey.
authors.forename: Lokman
authors.initials: L
authors.lastname: Ayaz
authors.affiliation: Department of Medicinal Biochemistry, Faculty of Medicine, Mersin University, Mersin, Turkey.
authors.forename: Lulufer
authors.initials: L
authors.lastname: Tamer Gumus
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mersin University, Mersin, Turkey.
authors.forename: Aysegul
authors.initials: A
authors.lastname: Gorur
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Yenisehir Campus, Mersin University, 33169 Mersin, Turkey. Electronic address: tunctanb@yahoo.com.
authors.forename: Bahar
authors.initials: B
authors.lastname: Tunctan
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D002374:Catalase / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D007249:Inflammation / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D048369:MAP Kinase Kinase 1 / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D018384:Oxidative Stress; D009195:Peroxidase / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000577:Amides; D011725:Pyridines; C108830:Y 27632; D030421:Peroxynitrous Acid; C002744:3-nitrotyrosine; D014443:Tyrosine; D008315:Malondialdehyde; D002374:Catalase; D009195:Peroxidase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D013482:Superoxide Dismutase; D019255:NADPH Oxidases; D054460:rho-Associated Kinases; D048369:MAP Kinase Kinase 1; D020742:rhoA GTP-Binding Protein; D005978:Glutathione
keywords: Ischemia/reperfusion; MEK1/ERK1/2/iNOS; Nitrosative stress; Oxidative stress; RhoA/Rho-kinase