title: Complement-dependent NADPH oxidase enzyme activation in renal ischemia/reperfusion injury.
abstract: NADPH oxidase plays a central role in mediating oxidative stress during heart, liver, and lung ischemia/reperfusion injury, but limited information is available about NADPH oxidase in renal ischemia/reperfusion injury. Our aim was to investigate the activation of NADPH oxidase in a swine model of renal ischemia/reperfusion damage. We induced renal ischemia/reperfusion in 10 pigs, treating 5 of them with human recombinant C1 inhibitor, and we collected kidney biopsies before ischemia and 15, 30, and 60 min after reperfusion. Ischemia/reperfusion induced a significant increase in NADPH oxidase 4 (NOX-4) expression at the tubular level, an upregulation of NOX-2 expression in infiltrating monocytes and myeloid dendritic cells, and 8-oxo-7,8-dihydro-2'-deoxyguanosine synthesis along with a marked upregulation of NADPH-dependent superoxide generation. This burden of oxidative stress was associated with an increase in tubular and interstitial expression of the myofibroblast marker α-smooth muscle actin (α-SMA). Interestingly, NOX-4 and NOX-2 expression and the overall NADPH oxidase activity as well as α-SMA expression and 8-oxo-7,8-dihydro-2'-deoxyguanosine synthesis were strongly reduced in C1-inhibitor-treated animals. In vitro, when we incubated tubular cells with the anaphylotoxin C3a, we observed an enhanced NADPH oxidase activity and α-SMA protein expression, which were both abolished by NOX-4 silencing. In conclusion, our findings suggest that NADPH oxidase is activated during ischemia/reperfusion in a complement-dependent manner and may play a potential role in the pathogenesis of progressive renal damage in this setting.
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Simone
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Medical and Surgical Sciences, University of Foggia, 71100 Foggia, Italy. Electronic address: federica.rascio@gmail.com.
authors.forename: F
authors.initials: F
authors.lastname: Rascio
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Castellano
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Divella
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Chieti
authors.affiliation: Urology, Andrology, and Renal Transplantation Unit, and Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Ditonno
authors.affiliation: Urology, Andrology, and Renal Transplantation Unit, and Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Battaglia
authors.affiliation: Veterinary Surgery Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Crovace
authors.affiliation: Veterinary Surgery Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Staffieri
authors.affiliation: Pharming BV, Leiden, The Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Oortwijn
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Medical and Surgical Sciences, University of Foggia, 71100 Foggia, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Stallone
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Gesualdo
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari "Aldo Moro," 70121 Bari, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Pertosa
authors.affiliation: Nephrology, Dialysis, and Transplantation Unit, Department of Medical and Surgical Sciences, University of Foggia, 71100 Foggia, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Grandaliano
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003174:Complement C1 Inactivator Proteins / Q000008:administration & dosage; D050718:Complement C1 Inhibitor Protein; D015926:Complement C3a / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D003713:Dendritic Cells / Q000502:physiology*; D003839:Deoxyadenosines / Q000096:biosynthesis / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D007684:Kidney Tubules / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D023421:Models, Animal; D019255:NADPH Oxidases / Q000378:metabolism*; D018384:Oxidative Stress; D034741:RNA, Small Interfering / Q000235:genetics; D015427:Reperfusion Injury / Q000201:enzymology* / Q000276:immunology; D034421:Sus scrofa
chemical_list: D000199:Actins; D003174:Complement C1 Inactivator Proteins; D050718:Complement C1 Inhibitor Protein; D003839:Deoxyadenosines; D034741:RNA, Small Interfering; C469952:SERPING1 protein, human; C078251:2'-deoxy-7,8-dihydro-8-oxoadenosine; D015926:Complement C3a; D003165:Complement System Proteins; D019255:NADPH Oxidases
keywords: C1 inhibitor; Complement system; Free radicals; NOX-2; NOX-4; Oxidative stress; Renal ischemia–reperfusion injury; Renal transplantation