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Physiome and pathophysiome research projects for studying cooperative functions of interstitial space and lymphatic system.

生理组和病理生理组研究项目，用于研究间质空间和淋巴系统的协作功能。

基本信息

批准号：
11470010
负责人：
OHHASHI Toshio
金额：
$ 1.47万
依托单位：
Shinshu University Graduate School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

We have designed and constructed a modified intravital microscope system for investigating murine lymphatic pump activity in vivo and developed a custom organ chamber with a semi-circular channel, being suitable for superfusion of murine mesentery in vivo. Using the new experimental appratus, we have evaluated whether or not thore is rhythmic pumping activity of murine mesenteric lymphatic vessels in vivo. A marked lymphatic pumping activity was observed in the mesenteries of DDY mice, The maximal and minimal diameter and frequency in the pumping activity were 60.9±1.0μm, 53.7±1.8μm and 12.8 min, respectively. Physiological roles of endogenous nitric oxide (NO) in the lymphatic pump activity in vivo were studied using the new appratus. A 15-min superfusion of 30μM-NAME in the mesenteries caused significant increasing the pumping frequency and the pump flow index and a significant decrease of the endo-diastoric and end-systolic diameters. Simultaneous superfusion of 1mM L-arginine with … More 30μM L-NAME produced a signficant reversal of the L-NAME-mediated increase of the frequency and decreace of the end-systolic diameter.We have attempted to examine whether the cultured lymphatic emdothelial cells, similar to angiogenesis of the blood vessels, have an ability to induce in vitro neovascularization of lymph vessels in response to basic fibroblast growth factor (bFGF). The effects of bFGF on the proliferation and migration of the cultured lymphatic endothelial cells isolated from dog thoracic ducts were evaluated by changing the number of the subconfluent cells and by wound migration assay, respectively. The effects of bFGF on invasion and tube formation of the cultured lymphatic endothelial cells into a three-dimensional collagen gel were also investigated by a phase-contrast microscope and an electron microscope. The 10ng/ml bFGF caused a significant induction of proferation and migration of the cultured lymphatic endothelial cells. The bFGF produced significantly invasion and tube formation of the cultured cells into a three-dimensional collagen gel. The bFGF also facilitated to form capillary-like tubes of the cultured cells between twe layers of collagen gets. These findings suggest that the cultured lymphatic endothelial cells have an ability to form lymphatic capillary-like tubes in response to bFGF. Less

我们设计并构建了一种改进的活体显微镜系统，用于研究小鼠体内淋巴泵活性，并开发了一种具有半圆形通道的定制器官室，适用于小鼠肠系膜的体内灌流。我们用这种新的实验装置对小鼠肠系膜淋巴管的节律性泵血活动进行了研究。DDY小鼠肠系膜有明显的淋巴泵活动，其最大、最小直径和频率分别为60.9±1.0μm、53.7±1.8μm和12.8min。利用该装置研究了内源性一氧化氮（NO）在体内淋巴泵活动中的生理作用。肠系膜灌注30μM-NAME 15 min，可使泵频率和泵流量指数显著增加，舒张末期内径和收缩末期内径显著减小。同时灌流1 mM L-精氨酸， ...更多信息 30μM的L-NAME可逆转L-NAME介导的收缩末期内径的增加和收缩末期内径的减小。分别通过改变亚融合细胞数量和伤口迁移实验来评价碱性成纤维细胞生长因子对犬胸导管淋巴管内皮细胞增殖和迁移的影响。用相差显微镜和电子显微镜观察bFGF对淋巴管内皮细胞三维胶原凝胶浸润和管腔形成的影响。10 ng/ml bFGF可明显诱导淋巴管内皮细胞增殖和迁移。bFGF可使培养的细胞向三维胶原凝胶中浸润并形成管状结构。bFGF还可促进培养细胞在两层胶原膜之间形成毛细血管样管。这些结果表明，培养的淋巴管内皮细胞有能力形成淋巴毛细血管样管的反应bFGF。少