The Role of CD14 and Toll-Like Receptor 4 of Kupffer Cells in Hepatic Ischemia-Reperfusion Injury in Rats
The Role of CD14 and Toll-Like Receptor 4 of Kupffer Cells in Hepatic Ischemia-Reperfusion Injury in Rats
复制标题
DOI：
10.1016/j.transproceed.2011.11.001
复制
发表时间：
2012-05-01
影响因子：
0.9
通讯作者：
Li, M.
中科院分区：
文献类型：
--
作者：
Luan, X.;Liu, Y.;Li, M.
objective.本研究旨在探讨枯否细胞（KCs）表面CD14和Toll样受体4（TLR4）在大鼠移植肝缺血再灌注损伤（IRI）中的作用。从对照组、IRI组和IRI+抗CD14抗体组获得分离的KC。我们测量了脂多糖受体CD 14和TLR 4的信使RNA（mRNA）和蛋白表达、核因子κ B（NF-κ β）活性和TNF-α水平。IRI组CD14和TLR4的mRNA和蛋白表达均显著高于对照组，流式细胞术和Western blot检测CD14和TLR4的蛋白表达也显著高于对照组。IRI组的NF-κ β活性和肿瘤坏死因子-α水平显著高于对照组（3.17 +/-0.21和0.28 +/-0.03 vs 654.2 +/-3.6 pg/mL和147.4 +/-1.1 pg/mL; t值分别为4.11和4.29; P <0.01）。与IRI组相比，在抗CD14抗体治疗后，它们显著降低：2.14 +/-0.17 vs 3.17 +/-0.21，298.7 +/-1.8 pg/mL vs 654.2 +/-3.6 pg/mL（t值分别= 2.52和2.92; P <0.05）。但仍显著高于对照组（t值分别为3.01和3.27; P <0.01）。IRI上调KC中CD14和TLR4基因表达，随后激活NF-κ β产生细胞因子。
Objective. 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.Methods. 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-kappa beta) activity, and TNF-alpha levels.Results. 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-kappa beta activity and tumor necrosis factor-alpha 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).Conclusion. IRI up-regulated CD14 and TLR4 gene expression in KCs, and subsequently activated NF-kappa beta to produce cytokines.