From the Department of Medicine, Division of Renal Diseases and Hypertension (H.N.H., J.P., M.C.M.W.-E., R.A.N.), Department of Pharmacology (A.O., M.C.M.W.-E., R.A.N.), and Cardiovascular and Pulmonary Research Laboratory (M.C.M.W.-E., R.A.N.), University of Colorado Denver, Aurora.
Arterioscler Thromb Vasc Biol. 2014 Apr;34(4):877-86. doi: 10.1161/ATVBAHA.114.303214. Epub 2014 Feb 13.
To define the contribution of vascular smooth muscle cell (SMC)-derived factors to macrophage phenotypic modulation in the setting of vascular injury.
By flow cytometry, macrophages (M4) were the predominant myeloid cell type recruited to wire-injured femoral arteries, in mouse, compared with neutrophils or eosinophils. Recruited macrophages from injured vessels exhibited a distinct expression profile relative to circulating mononuclear cells (peripheral blood monocytes; increased: interleukin-6, interleukin-10, interleukin-12b, CC chemokine receptor [CCR]3, CCR7, tumor necrosis factor-α, inducible nitric oxide synthase, arginase 1; decreased: interleukin-12a, matrix metalloproteinase [MMP]9). This phenotype was recapitulated in vitro by maturing rat bone marrow cells in the presence of macrophage-colony stimulating factor and 20% conditioned media from cultured rat SMC (sMϕ) compared with maturation in macrophage-colony stimulating factor alone (M0). Recombinant transforming growth factor (TGF)-β1 recapitulated the effect of SMC conditioned media. Macrophage maturation studies performed in the presence of a pan-TGF-β neutralizing antibody, a TGF-β receptor inhibitor, or conditioned media from TGF-β-depleted SMCs confirmed that the SMC-derived factor responsible for macrophage activation was TGF-β. Finally, the effect of SMC-mediated macrophage activation on SMC biology was assessed. SMCs cocultured with sMϕ exhibited increased rates of proliferation relative to SMCs cultured alone or with M0 macrophages.
SMC-derived TGF-β modulates the phenotype of maturing macrophages in vitro, recapitulating the phenotype found in vascular lesions in vivo. SMC-modulated macrophages induce SMC activation to a greater extent than control macrophages.
确定血管平滑肌细胞（SMC）衍生因子在血管损伤情况下对巨噬细胞表型调节的贡献。
通过流式细胞术，与中性粒细胞或嗜酸性粒细胞相比，在小鼠的线损伤股动脉中，巨噬细胞（M4）是募集的主要髓样细胞类型。与循环单核细胞（外周血单核细胞相比，来自损伤血管的募集巨噬细胞表现出明显不同的表达谱：白细胞介素-6、白细胞介素-10、白细胞介素-12b、CC 趋化因子受体 [CCR]3、CCR7、肿瘤坏死因子-α、诱导型一氧化氮合酶、精氨酸酶 1 增加；白细胞介素-12a、基质金属蛋白酶 [MMP]9 减少）。在存在巨噬细胞集落刺激因子和 20%培养的大鼠 SMC（sMϕ）条件培养基的情况下，体外培养大鼠骨髓细胞可重现这种表型，而在仅存在巨噬细胞集落刺激因子的情况下（M0）则不能。重组转化生长因子（TGF）-β1 重现了 SMC 条件培养基的作用。在存在泛 TGF-β 中和抗体、TGF-β 受体抑制剂或 TGF-β 耗尽的 SMC 条件培养基的情况下进行的巨噬细胞成熟研究证实，负责激活巨噬细胞的 SMC 衍生因子是 TGF-β。最后，评估了 SMC 介导的巨噬细胞激活对 SMC 生物学的影响。与单独培养的 SMC 或与 M0 巨噬细胞共培养的 SMC 相比，与 sMϕ 共培养的 SMC 表现出更高的增殖率。
SMC 衍生的 TGF-β 体外调节成熟巨噬细胞的表型，重现体内血管病变中发现的表型。与对照巨噬细胞相比，SMC 调节的巨噬细胞更能诱导 SMC 激活。
