[The influence with block the endotoxin signal transduction for ischemia/reperfusion injury of graft liver in rats].
OBJECTIVE
To explore the feasibility of interleukin 1 receptor associated kinase-4 (IRAK-4) as gene therapy target for liver ischemia/reperfusion injury (I/RI) and effective approach in vivo for short hairpin RNA (shRNA) interference used to gene therapy in liver graft hqappened.


METHODS
Sprague-Dawley rats were randomly divided into three groups: the control group, the in vivo transfection group (IVT) and the cold ischemia transfection group (CIT). Experiments of orthotopic liver transplantation were performed by two-cuff method. CIT were perfused with IRAK-4-shRNA plasmid (pSIIRAK-4) during cold ischemia phase, IVT received the equivalent volumes (2 mL) of pSIIRAK-4 after portal vein inosculated, and the control group leaved without any treatment. At 0 min, 60 min and 180 min after reperfusion, the expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot. The serum TNF-alpha level was detected by ELISA. Liver histopathological changes and cell apoptosis were observed by electron microscope and TUNEL.


RESULTS
After reperfusion, the expression of IRAK-4 were largely depressed in CIT than that of IVT and the control group (P < 0.01), and furthermore, the serum TNF-alpha level, proportion of hepatocyte apoptosis and severity of hepatocyte injury were also lower than the latter.


CONCLUSIONS
These results indicate that depression IRAK-4 expression with IRAK-4-shRNA through portal vein perfusion during cold ischemia phase could effectively blunt graft hepatic I/RI.
Liu
Zuo-jin
ZJ
Department of Hepatobiliary Surgery, the Second Clinical College, Chongqing University of Medical Sciences, Chongqing 400010, China.
Li
Sheng-wei
SW
Li
Xu-hong
XH
Peng
Yong
Y
You
Hai-bo
HB
Li
Shou-bai
SB
Gong
Jian-ping
JP
D000818:Animals; D015316:Genetic Therapy; D006084:Graft Rejection; D053592:Interleukin-1 Receptor-Associated Kinases / Q000096:biosynthesis / Q000235:genetics*; D008099:Liver / Q000098:blood supply* / Q000378:metabolism; D016031:Liver Transplantation / Q000009:adverse effects*; D008297:Male; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000235:genetics* / Q000517:prevention & control*; D015398:Signal Transduction; D014162:Transfection
D053592:Interleukin-1 Receptor-Associated Kinases