Butoi E, Gan A M, Tucureanu M M, Stan D, Macarie R D, Constantinescu C, Calin M, Simionescu M, Manduteanu I
Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania.
Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania; Laboratory of Molecular Medical Biochemistry, Nencki Institute of Experimental Bilogy, Warsaw, Poland.
Biochim Biophys Acta. 2016 Jul;1863(7 Pt A):1568-78. doi: 10.1016/j.bbamcr.2016.04.001. Epub 2016 Apr 7.
Coronary atherosclerosis complicated by plaque disruption and thrombosis is a critical event in myocardial infarction and stroke, the major causes of cardiovascular death. In atherogenesis, macrophages (MAC) and smooth muscle cells (SMC) are key actors; they synthesize matrix components and numerous factors involved in the process. Here, we design experiments to investigate whether SMC-MAC communication induces changes in ECM protein composition and/or neo-angiogenesis. Cell to cell communication was achieved using trans-well chambers, where SMCs were grown in the upper chamber and differentiated MAC in the bottom chamber for 24 or 72h. We found that cross-talk between MAC and SMC during co-culture: (i) significantly decreased the expression of ECM proteins (collagen I, III, elastin) in SMC; (ii) increased the expression and activity of metalloprotease MMP-9 and expression of collagenase MMP-1, in both MAC and SMC; (iii) augmented the secretion of soluble VEGF in the conditioned media of cell co-culture and VEGF gene expression in both cell types, compared with control cells. Moreover, the conditioned media collected from MAC-SMC co-culture promoted endothelial cell tube formation in Matrigel, signifying an increased angiogenic effect. In addition, the MAC-SMC communication led to an increase in inflammatory IL-1β and TLR-2, which could be responsible for cellular signaling. In conclusion, MAC-SMC communication affects factors and molecules that could alter ECM composition and neo-angiogenesis, features that could directly dictate the progression of atheroma towards the vulnerable plaque. Targeting the MAC-SMC cross-talk may represent a novel therapeutic strategy to slow-down or retard the plaque progression.
冠状动脉粥样硬化并发斑块破裂和血栓形成是心肌梗死和中风（心血管死亡的主要原因）中的关键事件。在动脉粥样硬化形成过程中，巨噬细胞（MAC）和平滑肌细胞（SMC）是关键因素；它们合成参与该过程的基质成分和众多因子。在此，我们设计实验来研究SMC-MAC通讯是否会诱导细胞外基质（ECM）蛋白组成变化和/或新生血管形成。使用Transwell小室实现细胞间通讯，其中SMC在上室生长，分化的MAC在下室生长24或72小时。我们发现共培养期间MAC和SMC之间的相互作用：（i）显著降低SMC中ECM蛋白（胶原蛋白I、III、弹性蛋白）的表达；（ii）增加MAC和SMC中金属蛋白酶MMP-9的表达和活性以及胶原酶MMP-1的表达；（iii）与对照细胞相比，细胞共培养条件培养基中可溶性血管内皮生长因子（VEGF）的分泌增加，且两种细胞类型中VEGF基因表达均增加。此外，从MAC-SMC共培养收集的条件培养基促进了基质胶中内皮细胞管形成，表明血管生成作用增强。此外，MAC-SMC通讯导致炎症因子IL-1β和Toll样受体2（TLR-2）增加，这可能负责细胞信号传导。总之，MAC-SMC通讯影响可能改变ECM组成和新生血管形成的因子和分子，这些特征可能直接决定动脉粥样硬化向易损斑块的进展。靶向MAC-SMC相互作用可能代表一种减缓或延缓斑块进展的新型治疗策略。
