Ravindran Dhanya, Ridiandries Anisyah, Vanags Laura Z, Henriquez Rodney, Cartland Siân, Tan Joanne T M, Bursill Christina A
Heart Research Institute, Newtown, Sydney, New South Wales, Australia.
Sydney Medical School, University of Sydney, Camperdown, Sydney, New South Wales, Australia.
PLoS One. 2017 Mar 10;12(3):e0173224. doi: 10.1371/journal.pone.0173224. eCollection 2017.
Chemokines are important in macrophage recruitment and the progression of atherosclerosis. The 'M3' chemokine binding protein inactivates key chemokines involved in atherosclerosis (e.g. CCL2, CCL5 and CX3CL1). We aimed to determine the effect of M3 on plaque development and composition. In vitro chemotaxis studies confirmed that M3 protein inhibited the activity of chemokines CCL2, CCL5 and CX3CL1 as primary human monocyte migration as well as CCR2-, CCR5- and CX3CR1-directed migration was attenuated by M3. In vivo, adenoviruses encoding M3 (AdM3) or green fluorescence protein (AdGFP; control) were infused systemically into apolipoprotein (apo)-E-/- mice. Two models of atherosclerosis development were used in which the rate of plaque progression was varied by diet including: (1) a 'rapid promotion' model (6-week high-fat-fed) and (2) a 'slow progression' model (12-week chow-fed). Plasma chemokine activity was suppressed in AdM3-infused mice as indicated by significantly less monocyte migration towards AdM3 mouse plasma ex vivo (29.56%, p = 0.014). In the 'slow progression' model AdM3 mice had reduced lesion area (45.3%, p = 0.035) and increased aortic smooth muscle cell α-actin expression (60.3%, p = 0.014). The reduction in lesion size could not be explained by changes in circulating inflammatory monocytes as they were higher in the AdM3 group. In the 'rapid promotion' model AdM3 mice had no changes in plaque size but reduced plaque macrophage content (46.8%, p = 0.006) and suppressed lipid deposition in thoracic aortas (66.9%, p<0.05). There was also a reduction in phosphorylated p65, the active subunit of NF-κb, in the aortas of AdM3 mice (37.3%, p<0.0001). M3 inhibited liver CCL2 concentrations in both models with no change in CCL5 or systemic chemokine levels. These findings show M3 causes varying effects on atherosclerosis progression and plaque composition depending on the rate of lesion progression. Overall, our studies support a promising role for chemokine inhibition with M3 for the treatment of atherosclerosis.
趋化因子在巨噬细胞募集和动脉粥样硬化进展过程中发挥着重要作用。“M3”趋化因子结合蛋白可使参与动脉粥样硬化的关键趋化因子失活（如CCL2、CCL5和CX3CL1）。我们旨在确定M3对斑块形成和组成的影响。体外趋化性研究证实，M3蛋白可抑制趋化因子CCL2、CCL5和CX3CL1的活性，因为M3可减弱原代人单核细胞的迁移以及CCR2、CCR5和CX3CR1介导的迁移。在体内，将编码M3的腺病毒（AdM3）或绿色荧光蛋白（AdGFP；对照）全身注入载脂蛋白（apo）-E-/-小鼠体内。采用了两种动脉粥样硬化发展模型，通过饮食改变斑块进展速度，包括：（1）“快速进展”模型（6周高脂喂养）和（2）“缓慢进展”模型（12周普通饲料喂养）。体外实验表明，与AdM3小鼠血浆共孵育时，单核细胞迁移显著减少（29.56%，p = 0.014），这表明AdM3注入小鼠体内后血浆趋化因子活性受到抑制。在“缓慢进展”模型中，AdM3小鼠的病变面积减小（45.3%，p = 0.035），主动脉平滑肌细胞α-肌动蛋白表达增加（60.3%，p = 0.014）。病变大小的减小无法用循环炎症单核细胞的变化来解释，因为AdM3组中的循环炎症单核细胞数量更高。在“快速进展”模型中，AdM3小鼠的斑块大小没有变化，但斑块巨噬细胞含量减少（46.8%，p = 0.006），胸主动脉脂质沉积受到抑制（66.9%，p<0.05）。AdM3小鼠主动脉中NF-κb的活性亚基磷酸化p65也减少（37.3%，p<0.0001）。在两种模型中，M3均抑制肝脏CCL2浓度，而CCL5或全身趋化因子水平没有变化。这些发现表明，根据病变进展速度，M3对动脉粥样硬化进展和斑块组成会产生不同影响。总体而言，我们的研究支持M3抑制趋化因子在治疗动脉粥样硬化方面具有广阔前景。
