Small interfering RNA targeting Fas protects mice against renal ischemia-reperfusion injury.
Fas-mediated apoptosis has been suggested to contribute to tubular cell death after renal ischemia-reperfusion injury. Here we investigate whether small interfering RNA (siRNA) duplexes targeting Fas protect mice from acute renal failure after clamping of the renal artery. Renal ischemia-reperfusion injury was induced by clamping the renal vein and artery for 15 or 35 min. Mice were treated before or after ischemia with siRNA targeting Fas or a control gene, administered by hydrodynamic injection, low-volume renal vein injection, or both. Treated mice were evaluated for renal Fas protein and mRNA expression, tissue histopathology, and apoptosis by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining. Blood urea nitrogen and survival were monitored in mice in which the contralateral kidney had been removed. A single hydrodynamic injection of Fas siRNA reduced Fas mRNA and protein expression in the kidney 4-fold. Kidneys from mice that received Fas siRNA two days earlier had substantially less renal tubular apoptosis by TUNEL staining and less tubular atrophy and hyaline damage. Whereas 90% of mice pretreated with saline or GFP siRNA died, only 20% of Fas-siRNA-pretreated animals died. The same survival advantage was provided by a single low-volume Fas siRNA injection into the renal vein. Moreover, postischemic injection through the renal vein protected 38% of mice from death. This study confirms the importance of Fas-mediated apoptosis in renal ischemia-reperfusion injury. Silencing Fas by systemic or local catheterization holds therapeutic promise to limit ischemia-reperfusion injury.
Institute of Pathophysiology, Semmelweis University, Nagyvárad tér 4, Budapest, H 1089, Hungary. hampet@net.sote.hu
Péter
P
Hamar
Erwei
E
Song
Gabor
G
Kökény
Allen
A
Chen
Nengtai
N
Ouyang
Judy
J
Lieberman
D000818:Animals; D001483:Base Sequence; D001806:Blood Urea Nitrogen; D020287:In Situ Nick-End Labeling; D007668:Kidney / Q000098:blood supply; D051379:Mice; D009838:Oligodeoxyribonucleotides; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D012079:Renal Circulation; D015427:Reperfusion Injury / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D019014:fas Receptor / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D034741:RNA, Small Interfering; D019014:fas Receptor