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Evaluation of the transport of micromolecular substances through Lymph node

小分子物质通过淋巴结转运的评估

基本信息

批准号：
14580842
负责人：
ONO Nobuyuki
金额：
$ 2.18万
依托单位：
Nagano National College of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580842/
关键词：
Lymph Lymph node Lymph transport Fluorescent tracer Intravital video microscope in-vivo リンパ循環流れ血管系可視化 VEGF 物質輸送透過性蛍光

项目摘要

The lymphatic system plays an important role in the regulating the transport of extracellular fluids and micromolecular substances in tissues and organs. The lymph travels between the regional lymph nodes. There is, however, few information of the flow profile within lymph nodes. In the present study, we have developed a fluorescent videomicroscopic system to visualize the flow profile within lymph nodes, and evaluate the lymph transport through the lymph nodes.The iliac lymph nodes were isolated from Wistar rats. The afferent and efferent lymph vessels of the isolated lymph nodes were cannulated in a domestic made organ chamber, and then the lymph nodes were selectively perfused with Krebs solution contained fluorescent tracers(FITC-dextran M.W. 71200, fluorescrite carboxylate microsphere φ 1μm and lymphocyte labeled with fluorescence). The flow images of the fluorescent tracers within the lymph node were obtained by a fluorescent videomicroscope via a SIT-camera and a chilled-CCD camera, and then the images were recorded to a video-recorder.The FITC-dextran perfused into the afferent lymph vessel of the lymph node, and then appeared to the efferent lymph vessel alter the perfusion at 30sec. On the other hand, the fluorescence carboxylate microsphere did not appear to the efferent lymph vessel after the perfusion at 20min. The lymphocyte labeled with fluorescence also did not appear to the efferent lymph vessel after the perfusion at 20min. The microspheres or lymphocytes accumulated in the lymph node. These results suggested that there are heterogeneities in the molecular size-dependent lymph flow pathway within the lymph node.

淋巴系统在调节细胞外液和小分子物质在组织器官中的转运中起着重要作用。淋巴液在区域淋巴结之间流动。然而，淋巴结内的流动轮廓的信息很少。在本研究中，我们开发了一个荧光视频显微镜系统，以显示淋巴结内的流动轮廓，并评估淋巴通过淋巴结的运输。将淋巴结的输入和输出淋巴管插入自制的器官腔中，然后用含荧光示踪剂的Krebs液（FITC-dextran M.W. 71200，荧光素微球φ 1μm，荧光标记淋巴细胞）。用荧光显微镜通过SIT摄像机和冷CCD摄像机获得淋巴结内荧光示踪剂的流动图像，并将图像记录在录像机中，FITC-葡聚糖灌注到淋巴结的输入淋巴管中，然后在灌注30秒后出现在输出淋巴管中。而荧光羧酸盐微球灌注20 min后，输出淋巴管未见荧光。灌注20 min后，输出淋巴管中也未见荧光标记的淋巴细胞。微球或淋巴细胞在淋巴结内聚集。这些结果表明，在淋巴结内的分子大小依赖的淋巴液流动途径中存在异质性。