Shao Zigong, Jiao Baoping, Yi Dehui, Liu Tingting, Pan Qi, Cheng Ying, Liu Hao
Department of General Surgery, the First Hospital of China Medical University, Shenyang, China.
Department of General Surgery, the First Hospital of Liaoning Medical University, Jinzhou, China.
Cell Physiol Biochem. 2017;41(4):1547-1554. doi: 10.1159/000470820. Epub 2017 Mar 27.
To investigate effect of warm ischemia after cardiac death on activation of TLR9 pathway in porcine liver.
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.
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.
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.
探讨心脏死亡后热缺血对猪肝中Toll样受体9（TLR9）信号通路激活的影响。
采用杜洛克、长白、大白三元杂交猪建立心脏死亡供体（DCD）模型。在热缺血后0、5、10、15、25和30分钟采集动物肝脏组织，分别通过实时荧光定量PCR和蛋白质印迹法分析TLR9、干扰素调节因子7（IRF7）、干扰素-β（IFN-β）和肿瘤坏死因子-α（TNF-α）在mRNA和蛋白质水平的表达，并通过蛋白质印迹法检测p65蛋白评估核因子κB（NF-κB）/DNA结合活性。
缺血以缺血时间依赖的方式诱导TLR9、IRF7、IFN-β和TNF-α在mRNA和蛋白质水平的表达。其中，TNF-α和IFN-β的表达诱导晚于TLR9和IRF7。缺血还增强了DCD肝脏组织中NF-κB与DNA的结合。用iCpG预处理可特异性阻断肝脏中缺血触发的TLR9信号通路激活，并保护动物免受缺血所致的肝组织损伤。
热缺血激活猪肝组织中的TLR9信号通路。阻断TLR9信号通路可为DCD肝移植中缺血所致的肝组织提供保护。
