Alleviation of ischemia-reperfusion injury in rat liver transplantation by induction of small interference RNA targeting Fas |
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Authors: | X Li J F Zhang M Q Lu Y Yang C Xu H Li G S Wang C J Cai G H Chen |
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Institution: | (1) Department of Liver Transplantation, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Road, Guangzhou, Guangdong Province, 510630, China |
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Abstract: | Background Cellular apoptosis plays an important role in ischemia-reperfusion (I/R) injury during organ transplantation. Synthetic small
interference RNA (siRNA) targeting apoptotic receptor Fas has proven effective to protect mice against hepatitis and renal
I/R injury. The objective of this study is to investigate the silencing impact of Fas siRNA to alleviate I/R injury in rat
liver transplantation.
Materials and methods Rat hepatocytes (BRL cells) were transfected with three pairs of synthesized Fas siRNA; cells untreated and treated with GFP
siRNA were taken as blank and siRNA control. The most effective Fas siRNA was chosen for in vivo experiments. Syngeneic orthotopic
liver transplantation was performed in Fas siRNA group, siRNA control group, and blank control group of Sprague–Dawley rats.
There were 25 pairs of rats in each group. siRNA transfection of donor rats was done with hydrodynamic injection method 48 h
before liver procurement. Blood and liver samples were collected for evaluation of serum ALT levels, Fas protein and mRNA
expression, and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, 1, 3,
6, 12, and 24 h after liver transplantation.
Results Fas siRNA2, which inhibited Fas gene expression much more than other siRNAs, was chosen for in vivo experiment. The serum
ALT levels of Fas siRNA group were much less than those of blank and siRNA control groups 1, 3, 6, 12, and 24 h after blood
reperfusion, indicating diminishing ischemia-reperfusion injury. Donor livers in Fas siRNA group had substantially less cell
apoptosis. The expression of Fas mRNA and protein was reduced dramatically in the Fas siRNA group compared with the other
two groups.
Conclusion Fas-mediated apoptosis play an important role in I/R injury of rat liver transplantation. Silencing Fas by hydrodynamic injection
of siRNA holds therapeutic promise to limit I/R injury. |
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Keywords: | Fas RNA interference Ischemia-reperfusion injury Syngeneic rat liver transplantation Apoptosis |
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