Interleukin 18 stimulates release of soluble lectin-like oxidized LDL receptor-1 (sLOX-1)
Interleukin 18 stimulates release of soluble lectin-like oxidized LDL receptor-1 (sLOX-1)
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DOI：
10.1016/j.atherosclerosis.2008.04.002
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发表时间：
2009-01-01
期刊：
影响因子：
5.3
通讯作者：
Kita, Toru
中科院分区：
文献类型：
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作者：
Mitsuoka, Hirokazu;Kume, Noriaki;Kita, Toru
凝集素样氧化 LDL 受体 1 (LOX-1) 似乎在动脉粥样硬化斑块破裂中发挥着至关重要的作用。我们之前报道过，急性冠脉综合征 (ACS) 中循环可溶性 LOX-1 (sLOX-1) 水平升高，并且 sLOX-1 可能是 ACS 的特异性且敏感的生物标志物。促炎细胞因子白细胞介素 18 (IL-18) 及其受体在动脉粥样硬化斑块中显着表达。此外，据报道 ACS 中循环 IL-18 水平较高。在这项研究中，我们检查了 IL-18 是否可以刺激 LOX-1 的脱落以及随后 sLOX-1 的释放。用LOX-1 cDNA转染后，HEK-293T细胞在有或没有IL-18的情况下孵育。使用抗 LOX-1 单克隆抗体对细胞条件培养基和总细胞裂解物进行免疫印迹分析。此外，还将ADAM10 cDNA、ADAM10 siRNA或对照载体共转染至HEK-293T细胞中，并在用或不用IL-18处理后对细胞条件培养基和总细胞裂解物进行LOX-1免疫印迹。将LOX-1或sLOX-1量的细胞条件培养基/总细胞裂解液的比率确定为sLOX-1裂解率。 IL-18 (10-100 ng/mL) 刺激以浓度和时间依赖性方式将 sLOX-1 裂解增加 3-4 倍。 ADAM10 单独过度表达同样增强了 sLOX-1 裂解。 ADAM10 siRNA 转染抑制 ADAM10 显着抑制 IL-18 诱导的 sLOX-1 裂解。 IL-18 同样增强了 TNF-α 激活的培养内皮细胞以及体内 LOX-1 转基因小鼠中 sLOX-1 的裂解。 IL-18 似乎是增强 ACS 中 sLOX-1 释放的刺激物之一，ADAM10 可能参与此过程。 (C) 2008 Elsevier Ireland Ltd. 保留所有权利。
Lectin-like oxidized LDL receptor-1 (LOX-1) appears to play crucial roles in atherosclerotic plaque rupture. We previously reported that circulating soluble LOX-1 (sLOX-1) levels are elevated in acute coronary syndrome (ACS) and that sLOX-1 can be a specific and sensitive biomarker for ACS. A proinflammatory cytokine interleukin 18 (IL-18) and its receptor are prominently expressed in atherosclerotic plaques. In addition, circulating IL-18 levels were reported to be high in ACS. In this study, we have examined if IL-18 can stimulate shedding of LOX-1 and subsequent release of sLOX-1. After transfection with LOX-1 cDNA, HEK-293T cells were incubated with or without IL-18. Cell-conditioned media and total cell lysates were subjected to immunoblot analyses with an anti-LOX-1 monoclonal antibody. In addition, ADAM10 cDNA, ADAM10 siRNA or control vector were also co-transfected into HEK-293T cells, and the cell-conditioned media and total cell lysates were subjected to LOX-1 immunoblotting after treatment with or without IL-18. The cell-conditioned medium/total cell lysate ratios in the amounts of LOX-1 or sLOX-1 were determined as sLOX-1 cleavage ratios. IL-18 (10-100 ng/mL) stimulation increased the sLOX-1 cleavage by 3-4-fold in a concentration- and time-dependent manner. ADAM10 over-expression alone similarly enhanced the sLOX-1 cleavage. ADAM10 inhibition by ADAM10 siRNA transfection significantly suppressed IL-18-induced sLOX-1 cleavage. IL-18 similarly enhanced sLOX-1 cleavage in TNF-alpha-activated cultured endothelial cells, as well as LOX-1 transgenic mice in vivo. IL-18 appears one of the stimuli that enhance sLOX-1 release in ACS and ADAM10 may be involved in this process. (C) 2008 Elsevier Ireland Ltd. All rights reserved.