Supplementary Material for: Differential Endothelial Coverage, Response to Injury and Neointimal Integration of CX3CR1/Smooth Muscle-Like Cells after Carotid or Femoral Arterial Injury

Background: Previously, we established the importance of the CX3CL1/CX3CR1 axis in the promotion of myeloid cell differentiation into neointimal smooth muscle-like cells (SMLC). Methods: In this study, acute (24 h) endothelial coverage and CX3CL1 expression as well as chronic (2 weeks) vascular remodeling was examined with respect to whether myeloid CX3CR1+ SMLC number in the neointima differed between carotid and femoral artery wire injury. Results and Conclusion: Twenty-four hours after injury, CX3CL1 expression was significantly elevated in injured carotid compared to femoral arteries. In mice with CX3CR1 promoter-driven expression of green fluorescent protein, neointima formation was significantly greater (p + cell integration was similar in both models, the carotid lesion had greater proportions of cells coexpressing CX3CR1 and both α-smooth muscle actin and calponin (p < 0.05). Wire injury of carotid arteries was associated with greater CX3CL1 expression in the acute phase followed by greater CX3CR1 coexpressing SMLC content in later lesions as well as less neointima formation than in femoral arteries. This may, in part, explain the variability in lesion composition after carotid versus femoral wire injury.背景：本团队既往已明确CX3CL1/CX3CR1轴在促进髓系细胞分化为新生内膜样平滑肌细胞（smooth muscle-like cells, SMLC）中的关键作用。方法：本研究针对颈动脉与股动脉钢丝损伤模型中，新生内膜内髓系CX3CR1阳性SMLC的数量是否存在差异，分别检测了急性期（24小时）的内皮覆盖情况、CX3CL1表达水平，以及慢性期（2周）的血管重塑情况。结果与结论：损伤后24小时，损伤侧颈动脉的CX3CL1表达水平较股动脉显著升高。在CX3CR1启动子驱动绿色荧光蛋白表达的小鼠中，颈动脉损伤组的新生内膜形成量显著高于股动脉损伤组（p < 0.05）。尽管两种模型的髓系细胞整合情况相似，但颈动脉损伤病灶中同时表达CX3CR1与α-平滑肌肌动蛋白及钙调蛋白的细胞占比更高（p < 0.05）。与股动脉钢丝损伤相比，颈动脉钢丝损伤在急性期伴随更高的CX3CL1表达，后期病灶中表达CX3CR1的SMLC含量也更高，而新生内膜形成量却更低。上述现象可在一定程度上解释颈动脉与股动脉钢丝损伤后病灶组成的差异。