Chang Ping, Liu Juan, Yu Ying, Cui Shao-Ye, Guo Zhen-Hui, Chen Gui-Ming, Huang Qiong, Liu Zhan-Guo
Department of Intensive Care Unit, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Geriatric Infection and Organ Function Support, Department of Medical Intensive Care Unit, General Hospital of Guangzhou Military Command, Guangzhou, China.
Cell Physiol Biochem. 2017;42(6):2559-2568. doi: 10.1159/000480217. Epub 2017 Aug 23.
BACKGROUND/AIMS: This study investigated signaling pathways via which extracellular histones induce the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) release from the macrophage cell line RAW 264.7 and the anti-inflammatory efficacy of the antioxidant alpha-lipoic acid (ALA).
ELISA and western blotting analyses were conducted to detect the release of TNF-α from histone-stimulated RAW 264.7 macrophages and the associated phospho-activation of MAPKs (ERK and p38) and NF-κB p65. The effects of ALA on the release of TNF-α and phospho-activation of the MAPKs and NF-κB p65 were studied. P < 0.05 was considered statistically significant.
Extracellular histones dose-dependently induced TNF-α release from RAW 264.7 cells and increased the phosphorylation of p38, ERK, and NF-κB p65. TNF-α release was markedly suppressed by p38, ERK, and NF-kB inhibitors. ALA reduced histone-induced TNF-α release, ERK/p38 MAPK activation, and NF-kB activation without affecting macrophage viability.
Histones induce TNF-α release from macrophages by activating the MAPK and NF-kB signaling pathways, while ALA suppresses this response by inhibiting ERK, p38 and NF-kB. These findings identify potentially critical inflammatory signaling pathways in sepsis and molecular targets for sepsis treatment.
背景/目的：本研究调查了细胞外组蛋白诱导巨噬细胞系RAW 264.7释放促炎细胞因子肿瘤坏死因子-α（TNF-α）的信号通路以及抗氧化剂α-硫辛酸（ALA）的抗炎功效。
进行酶联免疫吸附测定（ELISA）和蛋白质印迹分析，以检测组蛋白刺激的RAW 264.7巨噬细胞中TNF-α的释放以及丝裂原活化蛋白激酶（MAPKs，细胞外调节蛋白激酶ERK和p38）和核因子κB p65的相关磷酸化激活。研究了ALA对TNF-α释放以及MAPKs和核因子κB p65磷酸化激活的影响。P < 0.05被认为具有统计学意义。
细胞外组蛋白剂量依赖性地诱导RAW 264.7细胞释放TNF-α，并增加p38、ERK和核因子κB p65的磷酸化。p38、ERK和核因子κB抑制剂显著抑制TNF-α的释放。ALA可降低组蛋白诱导的TNF-α释放、ERK/p38丝裂原活化蛋白激酶激活和核因子κB激活，且不影响巨噬细胞活力。
组蛋白通过激活MAPK和核因子κB信号通路诱导巨噬细胞释放TNF-α，而ALA通过抑制ERK、p38和核因子κB来抑制这种反应。这些发现确定了脓毒症中潜在的关键炎症信号通路以及脓毒症治疗的分子靶点。
