Gene silencing of complement C5a receptor using siRNA for preventing ischemia/reperfusion injury.
Ischemia/reperfusion (I/R) injury in organ transplantation significantly contributes to graft failure and is untreatable using current approaches. I/R injury is associated with activation of the complement system, leading to the release of anaphylatoxins, such as C5a, and the formation of the membrane attack complex. Here, we report a novel therapy for kidney I/R injury through silencing of the C5a receptor (C5aR) gene using siRNA. Mice were injected with 50 microg of C5aR siRNA 2 days before induction of ischemia. Renal ischemia was then induced through clamping of the renal vein and artery of the left kidney for 25 minutes. The therapeutic effects of siRNA on I/R were evaluated by assessment of renal function, histopathology, and inflammatory cytokines. siRNA targeting C5aR efficiently inhibited C5aR gene expression both in vitro and in vivo. Administering C5aR siRNA to mice preserved renal function from I/R injury, as evidenced by reduced levels of serum creatinine and blood urea nitrogen in the treated groups. Inhibition of C5aR also diminished in vivo production of the pro-inflammatory cytokine tumor necrosis factor-alpha and chemokines MIP-2 and KC, resulting in the reduction of neutrophils influx and cell necrosis in renal tissues. This study demonstrates that siRNA administration represents a novel approach to preventing renal I/R injury and may be used in a variety of clinical settings, including transplantation and acute tubular necrosis.
Zheng
Xiufen
X
Department of Surgery, University of Western Ontario, London, Ontario, Canada.
Zhang
Xusheng
X
Feng
Biao
B
Sun
Hongtao
H
Suzuki
Motohiko
M
Ichim
Thomas
T
Kubo
Norihiko
N
Wong
Arthur
A
Min
Lisa R
LR
Budohn
Marianne E
ME
Garcia
Bertha
B
Jevnikar
Anthony M
AM
Min
Wei-Ping
WP
D000818:Animals; D001806:Blood Urea Nitrogen; D002460:Cell Line; D054426:Chemokine CXCL2 / Q000235:genetics / Q000378:metabolism; D003404:Creatinine / Q000097:blood; D015536:Down-Regulation; D020868:Gene Silencing; D007668:Kidney / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D007677:Kidney Function Tests; D008297:Male; D051379:Mice; D034741:RNA, Small Interfering / Q000378:metabolism*; D044087:Receptor, Anaphylatoxin C5a / Q000235:genetics*; D015427:Reperfusion Injury / Q000503:physiopathology / Q000517:prevention & control*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
D054426:Chemokine CXCL2; D034741:RNA, Small Interfering; D044087:Receptor, Anaphylatoxin C5a; D014409:Tumor Necrosis Factor-alpha; D003404:Creatinine
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