Immunohistochemical detection of intrathrombotic macrophage-derived cytokines and its application to thrombus age estimation in murine deep vein thrombosis model
Immunohistochemical detection of intrathrombotic macrophage-derived cytokines and its application to thrombus age estimation in murine deep vein thrombosis model
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DOI：
10.1007/s00414-013-0873-0
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发表时间：
2013-05
影响因子：
2.1
通讯作者：
M. Nosaka;Y. Ishida;A. Kimura;T. Kondo
中科院分区：
文献类型：
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作者：
M. Nosaka;Y. Ishida;A. Kimura;T. Kondo
我们用免疫化学方法检测了静脉血栓小鼠模型中IFNγ、TNFα和TNF受体p55（TNF-Rp 55）的表达。IFNγ+、TNFα+和TNF-Rp 55+细胞在下腔静脉结扎后7、3和3d开始出现。此后，这些阳性细胞的数量随着IVC结扎后的时间而增加。双色免疫荧光分析表明，每个分子表达的血栓内巨噬细胞。在所有结扎后间隔时间为7天或更短的样本中，IFNγ/巨噬细胞比率（IFNγ/巨噬细胞）、TNFα/巨噬细胞比率（TNFα/巨噬细胞）和TNF-Rp 55/巨噬细胞比率（TNF-Rp 55/巨噬细胞）分别<0.2、<0.2和<0.4。而IVC结扎后≥10 d，IFNγ/M β和TNFα/M β分别大于0.2和0.3。这些观察结果表明，IFNγ/M β比值>0.2和TNFα/M β比值>0.3表明血栓年龄≥10天。此外，TNF-Rp 55/M β比值>0.7强烈提示血栓年龄>14天。本研究表明，免疫组化法检测静脉血栓中IFNγ、TNFα和TNF-Rp 55的表达可用于判断血栓形成的年龄。然而，TNF-Rp 55/M比值的变化比其他两种标志物的变化更大。因此，IFNγ和TNFα可能是更实用的标志物。
We immunohistochemically examined the expression of IFNγ, TNFα, and TNF receptor p55 (TNF-Rp55) in a stasis-induced venous thrombus murine model. IFNγ+, TNFα+, and TNF-Rp55+cells could be first detected 7, 3, and 3 days after inferior vena cava (IVC) ligation. Thereafter, the numbers of these positive cells increased with time after IVC ligation. Double-color immunofluorescence analyses demonstrated that each molecule was expressed by intrathrombotic macrophages. In all samples with postligation intervals of 7 days or less, IFNγ/macrophage ratio (IFNγ/Mϕ), TNFα/macrophage ratio (TNFα/Mϕ), and TNF-Rp55/macrophage ratio (TNF-Rp55/Mϕ) were <0.2, <0.2, and <0.4, respectively. In contrast, IFNγ/Mϕ and TNFα/Mϕ were greater than 0.2 and 0.3 at ≥10 days after the IVC ligation, respectively. These observations suggested that IFNγ/Mϕ ratios of >0.2 and TNFα/Mϕ ratios of >0.3 indicated thrombus age of ≥10 days. Moreover, TNF-Rp55/Mϕ ratios of >0.7 strongly suggested thrombus age of >14 days. The present study demonstrated that the immunohistochemical detection of IFNγ, TNFα, and TNF-Rp55 was suitable to estimate the age of venous thrombi. However, the variation in TNF-Rp55/Mϕ ratios is wider than those of the other two markers. Thus, IFNγ and TNFα could be more practical markers.