Luan X, Liu Y, Li M
Department of Hepatobliliary Surgery, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing, China.
Transplant Proc. 2012 May;44(4):937-41. doi: 10.1016/j.transproceed.2011.11.001.
The objective of this study was to study the role of CD14 and Toll-like receptor 4 (TLR4) in Kupffer cells (KCs) on ischemia reperfusion injury (IRI) in rat liver grafts.
Isolated KCs were obtained from control, IRI, and IRI plus anti-CD14 antibody groups. We measured messenger RNA (mRNA) and protein expression of the lipopolysaccharide receptor CD14 and TLR4, nuclear factor kappa B (NF-κβ) activity, and TNF-α levels.
mRNA and protein expressions of CD14 and TLR4 were significantly higher in the IRI than in the control group, as were protein expressions of CD14 and TLR4 by flow cytometry and by Western blots. NF-κβ activity and tumor necrosis factor-α level in the IRI group were significantly higher than in the control group (3.17 ± 0.21 and 0.28 ± 0.03 vs 654.2 ± 3.6 pg/mL and 147.4 ± 1.1 pg/mL; t value = 4.11 and 4.29 for each; P < .01). Compared with the IRI group they were greatly decreased after anti-CD14 antibody treatment: 2.14 ± 0.17 vs 3.17 ± 0.21, 298.7 ± 1.8 pg/mL vs 654.2 ± 3.6 pg/mL (t value = 2.52 and 2.92 for each; P < .05). They were still significantly higher than the control group (t values of 3.01 and 3.27 for each; P < .01).
IRI up-regulated CD14 and TLR4 gene expression in KCs, and subsequently activated NF-κβ to produce cytokines.
本研究旨在探讨库普弗细胞（KCs）中CD14和Toll样受体4（TLR4）在大鼠肝移植缺血再灌注损伤（IRI）中的作用。
从对照组、IRI组和IRI加抗CD14抗体组获取分离的KCs。我们检测了脂多糖受体CD14和TLR4的信使核糖核酸（mRNA）和蛋白表达、核因子κB（NF-κβ）活性以及肿瘤坏死因子-α（TNF-α）水平。
IRI组中CD14和TLR4的mRNA和蛋白表达显著高于对照组，流式细胞术和蛋白质印迹法检测的CD14和TLR4蛋白表达也是如此。IRI组中的NF-κβ活性和肿瘤坏死因子-α水平显著高于对照组（分别为3.17±0.21和0.28±0.03，对比654.2±3.6 pg/mL和147.4±1.1 pg/mL；每组t值分别为4.11和4.29；P<.01）。与IRI组相比，抗CD14抗体治疗后它们显著降低：分别为2.14±0.17对比3.17±0.21，298.7±1.8 pg/mL对比654.2±3.6 pg/mL（每组t值分别为2.52和2.92；P<.05）。它们仍显著高于对照组（每组t值分别为3.01和3.27；P<.01）。
IRI上调了KCs中CD14和TLR4基因表达，随后激活NF-κβ以产生细胞因子。
