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ANALYSIS OF ENDOTHELIUM-DERIVED CONTRACTILE OR RELAXING FACTORS IN LYMPHATIC ENOTHELIAL CELLS

淋巴内皮细胞中内皮源性收缩或松弛因子的分析

基本信息

批准号：
04670047
负责人：
OHHASHI Toshio
金额：
$ 1.86万
依托单位：
SHINSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

Acetylcholine-induced release of nitric oxide from lymphatic endothelial cells : In precontracted dog thoracic ducts with endothelium, acetylcholine (ACh) produced dose-dependent relaxations. The ACh-induced relaxation was abolished by mechanical rubbing of the endothelium. Pretreatment with oxyhemoglobin, N^G-monomethy-L-arginine (L-NMMA) and methlene blue markedly supperssed the ACh-induced relaxation in the thoracic ducts with endothelium. ACh also caused a marked relaxation in the so-called sandwich-mounted preparation, demonstrating that the endothelial cells of the longitudinal strip in response to ACh must have released somo transferable substrance(s) that, on diffusion into the ring segment, activated relaxation of the precontracted lymphatic muscle cells. All these findings suggest that the ACh-induced relaxation is mainly through release of nitric oxide or its related compounds from the endothelial cells.Effects of ACh on spontaneous contractions in isolated bovine mesenteric … More limphatics : In isolated bovine mesenteric lymphatics with intact endothelium, ACh produced dose-dependent negative chronotropic and inotropic effects on the spontaneous contractions. The both effect induced by ACh were abolished by mechanical rubbing of the endothelium pretreatment with L-NMMA markedly supperessed the ACh-induced effects on spontaneous contractions in the lymph vessels. An additional treatment with L-arginine caused a marked reversal of the L-NMMA-induced inhibitory responses. The findings suggest that nitric oxide or its related compounds liberated from lymphatic endothelial cells seems to inhibit pacemaker activity of the spontaneous contractions and reduce the myogenic conduction and/or the mechanical activity of lymphatic smooth muscles.Tissue culture of lympatic endothelial cells in dog toracic ducts :The distal end of the lymph vessel was cannulated by a sterilized polyethylene catheter and then perfused with Rinaldini soultion containing 250 U/ml coolagenase and 0.5 mM EGTA.After 10 min incubation with collagenase, intraluminal space of the thoracic duct was perfused with minimum essential mediam containing 20 % fetal bovine serum, and then the perfusate was centrifuged to obtain the denuded lymphatic endothelial cells. The pellets of the cells were cultured at 37ﾟC in a humidified incubator with air and 5 % CO_2. The polygonal cells had a classic "cobblestone" morphology at confluence. Less

乙酰胆碱诱导淋巴管内皮细胞释放一氧化氮：在预收缩的狗胸导管内皮细胞中，乙酰胆碱（ACh）产生剂量依赖性舒张。机械摩擦内皮可阻断ACh诱导的血管舒张。氧合血红蛋白、N^G-单甲基-L-精氨酸（L-NMMA）和亚甲蓝预处理可明显抑制ACh对带内皮胸导管的舒张作用。ACh也引起所谓的环封标本的明显松弛，表明纵向条的内皮细胞对ACh的反应一定释放了一些可转移的物质，这些物质扩散到环段时，激活了预收缩的淋巴肌细胞的松弛。上述结果提示，ACh的舒张作用主要是通过内皮细胞释放一氧化氮或其相关化合物来实现的。ACh对离体牛肠系膜血管自发性收缩的影响 ...更多信息 limphatics：在内皮完整的离体牛肠系膜动脉，ACh对自发性收缩产生剂量依赖性的负性变时和变力作用。机械摩擦内皮可消除ACh的上述两种效应，L-NMMA预处理可显著抑制ACh引起的淋巴管自发收缩效应。用L-精氨酸的额外处理引起L-NMMA诱导的抑制反应的显著逆转。研究结果表明，从淋巴管内皮细胞释放的一氧化氮或其相关化合物似乎抑制了自发收缩的起搏活动，并减少了肌源性传导和/或淋巴管平滑肌的机械活动。用无菌聚乙烯导管插入淋巴管的远端，然后用含有250 U/ml coolagenase和0.5 mM EGTA的Rinaldini溶液灌注。用含有20%胎牛血清的最低必需培养基灌注胸导管的管腔内空间，然后离心灌注液以获得裸露的淋巴管内皮细胞。细胞团在37 ℃、空气和5%CO_2的加湿培养箱中培养.细胞融合时呈典型的“鹅卵石”状。少