Resin monomer-induced differential activation of MAP kinases and apoptosis in mouse macrophages and human pulp cells
Resin monomer-induced differential activation of MAP kinases and apoptosis in mouse macrophages and human pulp cells
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DOI：
10.1016/j.biomaterials.2010.01.005
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发表时间：
2010-04-01
期刊：
影响因子：
14
通讯作者：
Schweikl, Helmut
中科院分区：
文献类型：
--
作者：
Krifka, Stephanie;Petzel, Christine;Schweikl, Helmut
三甘醇二甲基丙烯酸酯（TEGDMA）是一种树脂单体，它是从聚合的牙科复合材料释放出来的。在长时间暴露后，它诱导各种靶细胞的凋亡或抑制LPS诱导的免疫系统细胞中细胞因子的产生。在这些组织中，丝裂原活化蛋白激酶（MAPK）调节支持细胞存活和细胞因子合成的信号转导途径。MAPK的时间依赖性调节以及它们与在树脂单体影响下诱导细胞凋亡和细胞因子释放的联系尚不清楚。本研究的目的是研究MAPK p38，JNK和ERK 1/2的上调或下调，诱导RAW264.7小鼠巨噬细胞和人牙髓衍生细胞的凋亡和细胞因子释放的动力学。在脂多糖（0.1 μ g/ml; LPS）（一种已知的MAPK活性诱导剂）和TEGDMA（3 mm）存在下，ERK 1/2、p38和JNK通过磷酸化被差异激活，如通过蛋白质印迹检测的。在巨噬细胞中，ERK 1/2被激活约6倍的LPS，而没有激活TEGDMA的存在下观察到15和30分钟后。一个轻微的激活p38检测到细胞培养后短时间暴露于TEGDMA（30分钟），但激活JNK后，确定只有LPS刺激。在长的24小时暴露期后，ERK 1/2和p38被LPS、LPS/TEGDMA的组合和单独的TEGDMA强烈激活（15-20倍）。在人牙髓来源的细胞中，ERK 1/2在暴露于TEGDMA长达2小时后被磷酸化，并且在长时间暴露24小时后检测到ERK 1/2以及p38的持续激活（1/2 -15倍）。LPS诱导的促炎细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）以及抗炎IL-10分泌的时间相关性增加在小鼠巨噬细胞中被TEGDMA瞬时抑制。同时，巨噬细胞培养物中处于凋亡和坏死阶段的细胞百分比随着暴露时间的增加而增加。然而，与细胞因子释放的抑制相反，LPS和TEGDMA引起的细胞凋亡和坏死在小鼠巨噬细胞和人牙髓源性细胞中都是晚期反应。从这些数据来看，似乎MAPK活化、细胞因子释放的抑制和TEGDMA诱导的细胞凋亡和坏死密切相关。然而，这些现象之间的直接因果关系需要进一步调查。(C)2010爱思唯尔有限公司版权所有。
Triethylene glycol dimethacrylate (TEGDMA) is a resin monomer which is released from polymerized dental composite materials. It induced apoptosis in various target cells or inhibition of LPS-induced cytokine production in cells of the immune system after prolonged exposure. In these tissues, mitogen-activated protein kinases (MAPK) regulate signal transduction pathways that support cell survival and cytokine synthesis. The time-dependent regulation of MAPK as well as their linkage to the induction of apoptosis and cytokine release under the influence of resin monomers is unknown. It was the aim of the present study to investigate the kinetics of the up- or down-regulation of the MAPK p38, JNK, and ERK1/2, the induction of apoptosis and cytokine release in RAW264.7 mouse macrophages and human pulp-derived cells. ERK1/2, p38 and JNK were differentially activated by phosphorylation in the presence of lipopolysaccharide (0.1 mu g/ml; LPS), a known inducer of MAPK activity, and TEGDMA (3 mm) as detected by Western blotting. In macrophages, ERK1/2 was activated about 6-fold by LPS, while no activation was observed in the presence of TEGDMA after 15 and 30 min. A slight activation of p38 was detected in cell cultures after short exposure to TEGDMA (30 min), but activated JNK was identified after LPS stimulation only. After a long 24 h exposure period, ERK1/2 and p38 were strongly activated by LPS, a combination of LPS/TEGDMA, and TEGDMA alone (15-20-fold). In human pulp-derived cells, ERK1/2 was phosphorylated after exposure to TEGDMA up to 2 h, and sustained activation of ERK1/2 as well as p38 (12-15-fold) was detected after prolonged exposure for 24 h. The LPS-induced, time-related increase in the secretion of the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) as well as the anti-inflammatory IL-10 was instantaneously inhibited by TEGDMA in mouse macrophages. In parallel, the percentage of cells in macrophage cultures in the stage of apoptosis and necrosis increased with exposure period. Yet, in contrast to the inhibition of cytokine release, apoptosis and necrosis caused by LPS and TEGDMA was a late response in both mouse macrophages and human pulp-derived cells. From these data it appears as if MAPK activation, inhibition of cytokine release and the induction of apoptosis and necrosis by TEGDMA are tightly related. The direct causal correlation of these phenomena, however, requires further investigation. (C) 2010 Elsevier Ltd. All rights reserved.