Interleukin-1 potently contributes to 25-hydroxycholesterol-induced synergistic cytokine production in smooth muscle cell-monocyte interactions
Interleukin-1 potently contributes to 25-hydroxycholesterol-induced synergistic cytokine production in smooth muscle cell-monocyte interactions
复制标题
DOI：
10.1016/j.atherosclerosis.2014.10.002
复制
发表时间：
2014-12-01
期刊：
影响因子：
5.3
通讯作者：
Loppnow, Harald
中科院分区：
文献类型：
--
作者：
Fu, Hang;Spieler, Florian;Loppnow, Harald
目的：炎症对于动脉粥样硬化形成至关重要。胆固醇是一种心血管危险因素，在此过程中可能会激活血管壁炎症。细胞因子介导的人类单核细胞与血管平滑肌细胞（SMC）的相互作用可能会延续这一过程。方法：我们研究了胆固醇代谢物 25-羟基胆固醇在新鲜分离的单核细胞与 SMC 共培养物中诱导炎症介质的能力。我们使用 qPCR、生物测定、ELISA 和蛋白质印迹确定了白细胞介素 (IL)-1 在这种相互作用中的作用。测试了 SMC 与单核细胞比例为 1:4 至 1:20 的共培养物。结果：在单独的 SMC 和单核细胞培养物（单一培养物）中，25-羟基胆固醇仅很少激活 IL-1、IL-6 和 MCP-1 的产生，而 LPS 刺激的细胞因子水平比未刺激的培养物高得多。相比之下，用 25-羟基胆固醇刺激的 SMC 和单核细胞的共培养物产生的细胞因子水平比相应的单一培养物高数百倍。 IL-1受体拮抗剂的阻断实验表明，IL-1对25-羟基胆固醇诱导的协同IL-6和MCP-1产生具有决定性作用。细胞内 IL-1 β 前体、释放的成熟 IL-1 β 和 caspase-1 p10 的存在表明炎症小体参与了这一过程。 Transwell 实验中 IL-1-mRNA 的测定表明单核细胞是 IL-1 的主要来源，IL-1 随后激活 SMC，而 SMC 是共培养物中 IL-6 的主要来源。结论：总而言之，局部血管壁细胞和入侵单核细胞之间的相互作用可能会增加血管壁中胆固醇引发的炎症，而 IL-1 可能在这一过程中发挥关键作用。数据还表明，低于单一培养物预期的胆固醇水平可能足以引发组织炎症。 (C) 2014 Elsevier Ireland Ltd. 保留所有权利。
Objectives: Inflammation is essential for atherogenesis. Cholesterol, a cardiovascular risk factor, may activate inflammation in the vessel wall during this process. Cytokine-mediated interactions of human monocytes with vascular smooth muscle cells (SMCs) may perpetuate this process. Methods: We investigated the capacity of the cholesterol metabolite 25-hydroxycholesterol to induce inflammatory mediators in cocultures of freshly isolated monocytes with SMCs. We determined the role of interleukin(IL)-1 in this interaction using qPCR, bioassays, ELISA and western blot. Cocultures with SMC to monocyte ratios from 1:4 to 1:20 were tested. Results: In separate SMC and monocyte cultures (monocultures) 25-hydroxycholesterol only poorly activated IL-1, IL-6 and MCP-1 production, whereas LPS stimulated much higher cytokine levels than unstimulated cultures. In contrast, cocultures of SMCs and monocytes stimulated with 25-hydroxycholesterol produced hundredfold higher cytokine levels than the corresponding monocultures. Blocking experiments with IL-1-receptor antagonist showed that IL-1 decisively contributed to the 25-hydroxycholesterol-induced synergistic IL-6 and MCP-1 production. The presence of intracellular IL-1 beta precursor, released mature IL-1 beta, and caspase-1 p10 indicated that the inflammasome was involved in this process. Determination of IL-1-mRNA in Transwell experiments indicated that the monocytes are the major source of IL-1, which subsequently activates the SMCs, the primary source of IL-6 in the coculture. Conclusion: Taken together, these interactions between local vessel wall cells and invading monocytes may multiply cholesterol-triggered inflammation in the vessel wall, and IL-1 may play a key role in this process. The data also indicate that lower cholesterol levels than expected from monocultures may suffice to initiate inflammation in the tissue. (C) 2014 Elsevier Ireland Ltd. All rights reserved.