Evaluation of the interaction between myogenic response and endothelium-derived factor by a microvessel perfusion system

通过微血管灌注系统评估生肌反应和内皮衍生因子之间的相互作用

• 批准号: 10555292
• 负责人: MOCHIZUKI Seiichi
• 金额: $ 6.34万
• 依托单位: Kawasaki College of Allied Health Professions
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10555292/
• 关键词: nitric oxide (NO); canine femoral artery; NO sensor; tetrahydrobiopterin; superoxide; fluorescent indicator; coronary circulation control; systemic NO production rate; 硝酸イオン; ラット摘出心; 潅流量; 蛍光計測; NADH; ラット腸間膜小動脈; ニトログリセリン; 微小電極

1. Isolated canine femoral artery : An NO microelectrode (100 μm in diameter) was inserted into the vascular media where flow-induced endothelium-derived NO production was measured NO production was changed with a time constant of about 24 sec, and a linear relation was observed between perfusion rate and produced NO.Flow-induced NO production was attenuated by exogenous NO (SNAP), and this suppressive effect was disappeared by perfusing BH_4, a cofactor of NO synthase (NOS). Simultaneous perfusion of SNAP and Tiron, superoxide scavenger did not affect endogenous NO production. Collectively, exogenous NO perfusion induces a decrease in intracellular BH_4 level, leading to superoxide release from NOS.Reaction between NO and superoxide is known to produce peroxynitrite, which attenuates NOS activity.2. Isolated rat mesenteric small artery : Intravascular-wall NO was visualized by an NO fluorescent indicator (DAF-2DA), and uniformly distributed NO fluorescence was observed during nitroglycerin (NTG) perfusion. In addition, it was found that thiol-containing molecules are involved in the NTG-derived NO production mechanism.3. Isolated perfused rat heart : When perfusion pressure was decreased from 100 to 50 cmH_2O, coronary perfusion rate decreased by half ; however, NO production was nearly doubled Despite decreased coronary perfusion rate by decreased perfusion pressure, NO production was increased in association with regulation mechanism of oxygen supply.4. Estimation method of systemic NO production rate : Plasma nitrate level in blood samples collected after simple 14-hr fasting was measured, and systemic NO production rate was estimated to be about 600 nmol/min by a single-compartment analysis. Compared with other previous estimation methods such as urinary nitrate and RI, estimated values were comparable.

1.犬离体股动脉：将直径为100μm的NO微电极插入血管中膜，测量血流诱导内皮源性NO的产生，NO产生的变化以约24秒的时间常数变化，并与灌流速率呈线性关系。外源性NO可减弱血流诱导的NO产生，而一氧化氮合酶的辅因子BH_4可使这种抑制作用消失。同时灌流超氧阴离子清除剂SNAP和TIron不影响内源性NO的产生。总体而言，外源性NO灌流诱导细胞内BH_4水平下降，导致NO释放超氧化物。已知NO与超氧化物反应产生过氧亚硝酸盐，从而减弱NOS的活性。分离的大鼠肠系膜小动脉：NO荧光指示剂(DAF-2DA)显示血管内壁NO，硝酸甘油(NTG)灌流过程中未见均匀分布的荧光。此外，还发现含有硫醇的分子参与了NTG衍生的NO的生成机制。大鼠离体心灌流：当灌流压力从100 cmH2O降至50cmH2O时，冠脉灌注率下降一半；而冠脉灌注率虽因灌流压降低而降低，但NO生成量增加近一倍，NO生成量增加与供氧调节机制有关。全身NO产生率的估算方法：测定单纯禁食14小时后采集的血样中硝酸盐水平，单室分析推算全身NO产生率约为600nmol/min。与以往的尿硝酸盐、RI等估算方法相比，估计值具有可比性。

项目成果

Seiichi Mochizuki: "Involvement of nitric oxide and oxidative stress in hemodialysis"Journal of Japanese Society of Biorheology. 14(4). 19-25 (2000)

望月精一：“血液透析中一氧化氮和氧化应激的参与”日本生物流变学会杂志。

Seiichi Mochizuki, et al.: "Suppression of flow-induced nitric oxide production by exogenous nitric oxide -involvement of tetrahydrobiopterin-"Journal of Japanese College of Angiology. 41(2). 117-121 (2001)

Seiichi Mochizuki等人：“外源性一氧化氮抑制血流诱导的一氧化氮生成-四氢生物蝶呤的参与-”日本血管学会杂志。

Mochizuki S et al.: "Flow dependence and time constant of the change in nitric oxide concentration measured in the vascular media"Medical & Biological Engineering & Computing. 37・4. 497-503 (1999)

Mochizuki S 等人：“血管介质中测量的一氧化氮浓度变化的流量依赖性和时间常数”医学与生物工程与计算 37・4（1999）。

Fumihiko Kajiya, et al.: "Biorheology in Biomimetics Handbook"N.T.S.. 1250 (2000)

Fumihiko Kajiya 等人：《仿生学手册中的生物流变学》N.T.S. 1250 (2000)

Mami Takemoto, et al.: "Intratissue NO measurement methods and their application"Technical Report of IEICE. MBE2000-50. 79-83 (2000)

Mami Takemoto等人：“组织内NO测量方法及其应用”IEICE技术报告。