Department of Anatomy and Embryology, George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, Romania.
Doctoral School of Medicine and Pharmacy, George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, Romania.
Int J Mol Sci. 2024 Mar 7;25(6):3097. doi: 10.3390/ijms25063097.
Monomeric C-reactive protein (mCRP) has recently been implicated in the abnormal vascular activation associated with development of atherosclerosis, but it may act more specifically through mechanisms perpetuating damaged vessel inflammation and subsequent aggregation and internalization of resident macrophages. Whilst the direct effects of mCRP on endothelial cells have been characterized, the interaction with blood monocytes has, to our knowledge, not been fully defined. Here we showed that mCRP caused a strong aggregation of both U937 cell line and primary peripheral blood monocytes (PBMs) obtained from healthy donors. Moreover, this increase in clustering was dependent on focal adhesion kinase (FAK) activation (blocked by a specific inhibitor), as was the concomitant adhesive attachment to the plate, which was suggestive of macrophage differentiation. Confocal microscopy confirmed the increased expression and nuclear localization of p-FAK, and cell surface marker expression associated with M1 macrophage polarization (CD11b, CD14, and CD80, as well as iNOS) in the presence of mCRP. Inclusion of a specific CRP dissociation/mCRP inhibitor (C10M) effectively inhibited PBMs clustering, as well as abrogating p-FAK expression, and partially reduced the expression of markers associated with M1 macrophage differentiation. mCRP also increased the secretion of pro-inflammatory cytokines Interleukin-8 (IL-8) and Interleukin-1β (IL-1β), without notably affecting MAP kinase signaling pathways; inclusion of C10M did not perturb or modify these effects. In conclusion, mCRP modulates PBMs through a mechanism that involves FAK and results in cell clustering and adhesion concomitant with changes consistent with M1 phenotypical polarization. C10M has potential therapeutic utility in blocking the primary interaction of mCRP with the cells-for example, by protecting against monocyte accumulation and residence at damaged vessels that may be predisposed to plaque development and atherosclerosis.
单体 C 反应蛋白（mCRP）最近被牵连到与动脉粥样硬化发展相关的异常血管激活中，但它可能通过更具体的机制发挥作用，这些机制会使受损血管炎症持续存在，并随后使驻留巨噬细胞聚集和内化。虽然已经对 mCRP 对内皮细胞的直接作用进行了描述，但据我们所知，其与血液单核细胞的相互作用尚未完全定义。在这里，我们表明 mCRP 可强烈聚集 U937 细胞系和来自健康供体的外周血单核细胞（PBM）。此外，这种聚集的增加依赖于粘着斑激酶（FAK）的激活（被特异性抑制剂阻断），同时伴随着对平板的附着，这表明发生了巨噬细胞分化。共聚焦显微镜证实，在 mCRP 存在的情况下，p-FAK 的表达增加和核定位增加，以及与 M1 巨噬细胞极化相关的细胞表面标志物（CD11b、CD14 和 CD80 以及 iNOS）的表达增加。包含特异性 CRP 解离/mCRP 抑制剂（C10M）可有效抑制 PBM 聚集，以及阻断 p-FAK 的表达，并部分降低与 M1 巨噬细胞分化相关的标志物的表达。mCRP 还增加了促炎细胞因子白细胞介素-8（IL-8）和白细胞介素-1β（IL-1β）的分泌，而不会显著影响 MAP 激酶信号通路；包含 C10M 不会扰乱或改变这些作用。总之，mCRP 通过涉及 FAK 的机制调节 PBM，导致细胞聚集和附着，同时伴随着与 M1 表型极化一致的变化。C10M 具有阻断 mCRP 与细胞的初次相互作用的潜在治疗用途，例如通过防止单核细胞在受损血管中积累和驻留，这些血管可能容易发生斑块形成和动脉粥样硬化。
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