Prevention of TLR9 Pathway in Warm Ischemia in Porcine Donor Liver after Cardiac Death.
OBJECTIVE
To investigate effect of warm ischemia after cardiac death on activation of TLR9 pathway in porcine liver.


METHODS
Donor of cardiac death (DCD) model was established with Duroc, Landrace, Large White crossbred pigs. Liver tissues from the animals were harvested at 0, 5, 10, 15, 25 and 30 minutes after warm ischemia for analysis of expression of TLR9, IRF7, IFN-β, and TNF-α at mRNA and protein levels by real-time PCR and western blot, respectively, and for assessment of NF-κB/DNA binding activity by western blot detection of p65 protein.


RESULTS
Ischemia induced TLR9, IRF7, IFN-β, and TNF-α expression at both mRNA and protein levels in an ischemic time dependent manner. Among them, expression of TNF-α and IFN- β was induced later than TLR9 and IRF7 did. Ischemia also enhanced NF-κB binding to DNA in the DCD liver tissue. Pretreatment with iCpG specifically blocked activation of TLR9 pathway triggered by ischemia in liver and protected the animals from ischemia-caused liver tissue damage.


CONCLUSIONS
Warm ischemia activates TLR9 pathways in the porcine liver tissue. Blocking TLR9 pathway could offer protection from ischemia-caused liver tissue in DCD liver transplantation.
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Zigong
Z
Shao
Department of General Surgery, the First Hospital of Liaoning Medical University, Jinzhou, China.
Baoping
B
Jiao
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Dehui
D
Yi
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Tingting
T
Liu
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Qi
Q
Pan
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Ying
Y
Cheng
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Hao
H
Liu
D000818:Animals; D003643:Death; D005786:Gene Expression Regulation; D050839:Interferon Regulatory Factor-7 / Q000096:biosynthesis; D016899:Interferon-beta / Q000096:biosynthesis; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D015398:Signal Transduction; D013552:Swine; D051217:Toll-Like Receptor 9 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D052096:Warm Ischemia
D050839:Interferon Regulatory Factor-7; D051217:Toll-Like Receptor 9; D014409:Tumor Necrosis Factor-alpha; D016899:Interferon-beta
Ischemia; Liver transplant; Signal transduction; Toll-like receptor