Immunohistochemical detection of intrathrombotic fibrocytes and its application to thrombus age estimation in murine deep vein thrombosis model
Immunohistochemical detection of intrathrombotic fibrocytes and its application to thrombus age estimation in murine deep vein thrombosis model
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DOI：
10.1007/s00414-016-1465-6
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发表时间：
2017-01-01
影响因子：
2.1
通讯作者：
Kondo, Toshikazu
中科院分区：
文献类型：
--
作者：
Nosaka, Mizuho;Ishida, Yuko;Kondo, Toshikazu
随着血栓年龄的增长，血栓在纤维化过程中沿着溶解。纤维细胞是具有间充质特征的循环骨髓源性细胞，其在纤维化中可能具有独特和关键的功能。在这项研究中，进行了双色免疫荧光分析，通过使用抗CD45和抗胶原I型抗体，以检查时间依赖性的外观的纤维细胞在小鼠模型的淤滞诱导的深静脉血栓形成。血栓年龄为1、3、5、7、10、14和21天。在血栓年龄小于5天，从未检测到CD45（+）和I型胶原+纤维细胞。血栓内纤维细胞最初见于7天龄的血栓，随着血栓年龄的增长，其数量逐渐增多。在定量形态计量学分析中，血栓内纤维细胞的平均数量在14天龄的血栓中最高，并且所有14天龄的5个样品的纤维细胞数量均大于25，其中3个样品的血栓内纤维细胞数量大于30。相反，在所有血栓样本中，结扎后间隔10和21天，血栓内纤维细胞的数量小于25。这些观察结果表明，含有纤维细胞的血栓至少存在7天，纤维细胞数量超过30表明血栓存在约14天。我们的观察表明，纤维细胞的检测可能是有用的血栓年龄测定。
Deep vein thrombi are dissolved after fibrosis process along with an increase of thrombus age. Fibrocytes are circulating bone marrow-derived cells with mesenchymal features that potentially have a unique and critical function in fibrosis. In this study, a double-color immunofluorescence analysis was carried out by using anti-CD45 and anti-collagen type I antibodies to examine the time-dependent appearance of fibrocytes in the murine model of stasis-induced deep vein thrombosis. The thrombus ages were 1, 3, 5, 7, 10, 14, and 21 days. In a thrombus age of less than 5 days, CD45(+) and collagen type I+ fibrocytes were never detected. The intrathrombotic fibrocytes were initially observed in thrombi aged 7 days, and their number increased with advances in thrombus age. In a quantitative morphometrical analysis, the average number of intrathrombotic fibrocytes was highest in 14-day-old thrombi, and all of the five samples aged 14 days had the fibrocyte number of more than 25, and in three out of them, the number of intrathrombotic fibrocytes was over 30. On the contrary, in all of thrombus samples with the postligation intervals of 10 and 21 days, the number of intrathrombotic fibrocytes was less than 25. These observations imply that thrombi containing fibrocytes are at least 7 days old and that a fibrocyte number exceeding 30 would indicate the thrombus age of approximately 14 days. Our observations indicate that the detection of fibrocytes could be useful for thrombus age determination.