Majkova Zuzana, Smart Eric, Toborek Michal, Hennig Bernhard
Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536-0200, USA.
Toxicol Appl Pharmacol. 2009 May 15;237(1):1-7. doi: 10.1016/j.taap.2009.02.016. Epub 2009 Mar 2.
Atherosclerosis, the primary cause of heart disease and stroke is initiated in the vascular endothelium, and risk factors for its development include environmental exposure to persistent organic pollutants. Caveolae are membrane microdomains involved in regulation of many signaling pathways, and in particular in endothelial cells. We tested the hypothesis that intact caveolae are required for coplanar PCB77-induced up-regulation of monocyte chemoattractant protein-1 (MCP-1), an endothelium-derived chemokine that attracts monocytes into sub-endothelial space in early stages of the atherosclerosis development. Atherosclerosis-prone LDL-R(-/-) mice (control) or caveolin-1(-/-)/LDL-R(-/-) mice were treated with PCB77. PCB77 induced aortic mRNA expression and plasma protein levels of MCP-1 in control, but not caveolin-1(-/-)/LDL-R(-/-) mice. To study the mechanism of this effect, primary endothelial cells were used. PCB77 increased MCP-1 levels in endothelial cells in a time- and concentration-dependent manner. This effect was abolished by caveolin-1 silencing using siRNA. Also, MCP-1 up-regulation by PCB77 was prevented by inhibiting p38 and c-Jun N-terminal kinase (JNK), but not ERK1/2, suggesting regulatory functions via p38 and JNK MAPK pathways. Finally, pre-treatment of endothelial cells with the aryl hydrocarbon receptor (AhR) inhibitor alpha-naphthoflavone (alpha-NF) partially blocked MCP-1 up-regulation. Thus, our data demonstrate that coplanar PCB77 can induce MCP-1 expression by endothelial cells and that this effect is mediated by AhR, as well as p 38 and JNK MAPK pathways. Intact caveolae are required for these processes both in vivo and in vitro. This further supports a key role for caveolae in vascular inflammation induced by persistent organic pollutants.
动脉粥样硬化是心脏病和中风的主要病因,始于血管内皮,其发展的风险因素包括环境中接触持久性有机污染物。小窝是参与多种信号通路调节的膜微区,尤其是在内皮细胞中。我们检验了这样一个假设,即完整的小窝对于共平面多氯联苯77(PCB77)诱导单核细胞趋化蛋白-1(MCP-1)的上调是必需的,MCP-1是一种内皮细胞源性趋化因子,在动脉粥样硬化发展的早期阶段吸引单核细胞进入内皮下空间。将易患动脉粥样硬化的低密度脂蛋白受体敲除小鼠(对照组)或小窝蛋白-1敲除/低密度脂蛋白受体敲除小鼠用PCB77处理。PCB77诱导对照组小鼠主动脉中MCP-1的mRNA表达和血浆蛋白水平升高,但在小窝蛋白-1敲除/低密度脂蛋白受体敲除小鼠中未出现这种情况。为了研究这种效应的机制,使用了原代内皮细胞。PCB77以时间和浓度依赖性方式增加内皮细胞中MCP-1的水平。使用小干扰RNA沉默小窝蛋白-1可消除这种效应。此外,抑制p38和c-Jun氨基末端激酶(JNK)可阻止PCB77对MCP-1的上调作用,但抑制细胞外信号调节激酶1/2(ERK1/2)则无此作用,这表明通过p38和JNK丝裂原活化蛋白激酶(MAPK)途径发挥调节功能。最后,用芳烃受体(AhR)抑制剂α-萘黄酮(α-NF)预处理内皮细胞可部分阻断MCP-1的上调。因此,我们的数据表明,共平面PCB77可诱导内皮细胞表达MCP-1,且这种效应由AhR以及p38和JNK MAPK途径介导。完整的小窝在体内和体外的这些过程中都是必需的。这进一步支持了小窝在持久性有机污染物诱导的血管炎症中起关键作用。