title: Effect of Adenoviral Catalase Gene Transfer on Renal Ischemia/Reperfusion Injury in Rats.
abstract: Ischemia/reperfusion (I/R) may through overt H₂O₂-induced pathophysiologic mechanisms lead to renal dysfunction. We explore whether catalase (CAT) protein overexpression by adenoviral CAT gene (Adv-CAT) transfection may improve ischemia/reperfusion-induced renal dysfunction. We augmented renal CAT expression by intrarenal arterial Adv-CAT administration with renal venous clamping in avertin-anesthetized female Wistar rats. After Adv-CAT transfection, we examined the CAT expression, location and effects on blood urea nitrogen (BUN) and urinary tubular injury biomarkers by biochemical assays, microcirculation by a laser perfusion imager, renal H₂O₂ amount by a chemiluminescent analyzer and molecular mechanisms including cytosolic cytochrome C leakage, apoptosis, autophagy and phospho- Akt (p-Akt)/phospho-endothelial nitric oxide (p-eNOS)/nitric oxide (NO) signaling by western blotting, immunohistochemistry and immunofluorescence. Adv-CAT enhanced 2.6-fold renal CAT protein expression primarily located in the proximal and distal tubules and renal vessels. Ischemia/reperfusion increased cytosolic cytochrome C leakage, renal H₂O₂-dependent level, autophagic Beclin-1/Atg5-Atg12/LC3 II expression, apoptotic Bax/Bcl-2/caspase 3/poly-(ADP-ribose)-polymerase fragments (PARP) expression and terminal deoxynucleotidyl transferasemediated nick-end labeling (TUNEL) stains and BUN and urinary glutathione S-transferase (GST) levels leading to proximal tubular injury. Ischemia/reperfusion also decreased renal microvascular blood flow associated with the inhibited renal expression of p-Akt and p-eNOS and NO production. Adv-CAT significantly improved the reduction in renal microvascular blood flow, reduced ischemia/reperfusion-enhanced oxidative stress, Beclin-1/Atg5-Atg12/LC3 II-meidated autophagy, Bax/Bcl-2/caspase 3/PARP-mediated apoptotic signaling, TUNEL stains, urinary GST level, and restored the p-Akt/p-eNOS/NO signaling in the kidney. Treatment of phosphatidylinositol 3-kinase inhibitors, wortmannin and LY294002, deleted Adv-CAT-induced p-Akt/p-eNOS/NO protective signaling. In conclusion, our results suggest Adv-CAT gene transfer counteracts H₂O₂-induced ischemia/reperfusion injury through preserving p-Akt/p-eNOS/NO pathway in the rat kidney.
authors.affiliation: Department of Internal Medicine, National Yang-Ming University, Taipei 11221, Taiwan, Republic of China.|Department of Planning, Ministry of Health and Welfare, Executive Yuan, Taipei 11558, Taiwan, Republic of China.
authors.forename: Chih-Ching
authors.initials: CC
authors.lastname: Yang
authors.affiliation: Department of Internal Medicine, Far-Eastern Memorial Hospital, New Taipei City 22056, Taiwan, Republic of China.
authors.forename: Ship-Ping
authors.initials: SP
authors.lastname: Hsu
authors.affiliation: Division of General Surgery, Far-Eastern Memorial Hospital, New Taipei City 22056, Taiwan, Republic of China.
authors.forename: Kuo-Hsin
authors.initials: KH
authors.lastname: Chen
authors.affiliation: Departments of Life Science, National Taiwan Normal University, Taipei 11677, Taiwan, Republic of China.
authors.forename: Chiang-Ting
authors.initials: CT
authors.lastname: Chien
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002374:Catalase / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy; D007668:Kidney / Q000098:blood supply*; D009569:Nitric Oxide / Q000502:physiology; D052250:Nitric Oxide Synthase Type III / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000628:therapy*
chemical_list: D009569:Nitric Oxide; D002374:Catalase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D051057:Proto-Oncogene Proteins c-akt