title: Protection of renal ischemia injury using combination gene silencing of complement 3 and caspase 3 genes.
abstract: BACKGROUND
  Ischemia/reperfusion (I/R) injury occurs in clinical kidney transplantation, which results in graft dysfunction and rejection. It has been documented that I/R injury is associated with complement activation and renal cell apoptosis. The purpose of this study was to develop a strategy to prevent I/R injury using small interfering RNA (siRNA) that target complement 3 (C3) and caspase 3 genes.
  
  
  METHODS
  siRNA-expression vectors were constructed to target C3 and caspase 3 genes. Gene silencing efficacy was assessed using real-time polymerase chain reaction. In vivo gene silencing was performed by hydrodynamic injection with C3 and caspase 3 siRNA. Renal I/R injury was induced through clamping the renal vein and artery for 25 min. I/R injury was evaluated using kidney histopathology, blood urea nitrogen (BUN), serum levels of creatinine, and survival.
  
  
  RESULTS
  Effective gene silencing was first confirmed in vitro. Notably upregulated expression of C3 and caspase 3 genes was observed from 2 to 48 hr after I/R injury, which were effectively and specifically inhibited by C3 and caspase 3 siRNA. In comparison with control mice, serum levels of creatinine and BUN were also significantly decreased in C3 and caspase 3 siRNA-treated mice. Furthermore, the therapeutic effect of siRNA was assessed in a severe, lethal I/R injury experiment, in which siRNA treatment significantly reduced mortality. Tissue histopathology showed an overall reduction in injury area in siRNA-treated mice.
  
  
  CONCLUSIONS
  This is the first demonstration that renal I/R injury can be prevented through silencing the complement gene and apoptosis gene, highlighting the potential for siRNA-based clinical therapy.
authors.affiliation: Department of Surgery, University of Western Ontario, London, Ontario, Canada.
authors.forename: Xiufen
authors.initials: X
authors.lastname: Zheng
authors.forename: Xusheng
authors.initials: X
authors.lastname: Zhang
authors.forename: Hongtao
authors.initials: H
authors.lastname: Sun
authors.forename: Biao
authors.initials: B
authors.lastname: Feng
authors.forename: Mu
authors.initials: M
authors.lastname: Li
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.forename: Costin
authors.initials: C
authors.lastname: Vladau
authors.forename: Dong
authors.initials: D
authors.lastname: Chen
authors.forename: Motohiko
authors.initials: M
authors.lastname: Suzuki
authors.forename: Lisa
authors.initials: L
authors.lastname: Min
authors.forename: Weihua
authors.initials: W
authors.lastname: Liu
authors.forename: Robert
authors.initials: R
authors.lastname: Zhong
authors.forename: Bertha
authors.initials: B
authors.lastname: Garcia
authors.forename: Anthony
authors.initials: A
authors.lastname: Jevnikar
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Min
mesh_terms: D000818:Animals; D053148:Caspase 3 / Q000235:genetics; D061945:Caspase Inhibitors; D002478:Cells, Cultured; D003176:Complement C3 / Q000037:antagonists & inhibitors* / Q000235:genetics; D015316:Genetic Therapy; D007668:Kidney / Q000098:blood supply* / Q000473:pathology; D016030:Kidney Transplantation; D051379:Mice; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000627:therapeutic use; D015427:Reperfusion Injury / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*
chemical_list: D061945:Caspase Inhibitors; D003176:Complement C3; D034741:RNA, Small Interfering; D053148:Caspase 3