Analysis of abnormal coronary micro-circulation in a diabetic or hypertensive rat heart by simultaneous measurement of NADH fluorescence and blood flow distribution

同时测量NADH荧光和血流分布分析糖尿病或高血压大鼠心脏冠状动脉微循环异常

• 批准号: 11680856
• 负责人: NAKAMOTO Hiroshi
• 金额: $ 2.56万
• 依托单位: Kawasaki Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680856/
• 关键词: coronary microcirculation; diabetes mellitus; hypertension; NADH; molecular tracer; heterogeneity; ischaemia; 血流分子トレーサ; 蛍光計測用CCDカメラ

There are two features of our study. Direct visualization of mitochondrial hypoxic state of the myocytes by nicotinamide adenine dinucleotide (NADH) fluorescence and blood flow visualization with molecular flow tracers with high resolution, 40 microns and 100 microns respectively. We mearsured NADH fluorescence intensity excited by ultraviolet ray in Langendorff preparation of diabetic rats with or without ischaemic condition. We measured blood flow distribution by perfusing tritiated desmethyl-imipramine (^3H-DMI) into the myocardium, which was later sliced and printed on an imaging plate. There is not any precedent study that measured at a time mitochondrial oxygen metabolism and myocardial blood flow distribution, which are most significant to coronary microcirculation. We also dealt with hypertensive rats in our study.From the NADH fluorescence study, we found that there is blood flow distribution heterogeneity in myocardial microcirculation, which is enhanced during ischaemia. This was confirmed by the molecular tracer method. The minimum unit length of microcirculation was in the same size in both methods, about 400 microns. As to basic research on hypertension, asymmetric dimethylarginine (ADMA) level was significantly elevated in relation to nitric oxide (NO) production compared with control rats. In NADH fluorescence study of diabetic rats, the time constant from normoxic to hypoxic state tended to be shorter and the time constant from hypoxic to normoxic state tended to be longer. This supports that diabetic rat hearts are vulnerable to ischaemia. In addition, reperfusion blood flow after ischaemic condition is deeply related to NO production.

我们的研究有两个特点。采用烟酰胺腺嘌呤二核苷酸（nictinamide adenine dinucleotide， NADH）荧光直接显示肌细胞线粒体缺氧状态，采用高分辨率40微米和100微米的分子流示踪剂直接显示血流。用紫外激发法测定了缺血性和非缺血性糖尿病大鼠Langendorff制剂中NADH的荧光强度。我们通过向心肌灌注氚化去甲基丙咪嗪（^3H-DMI）来测量血流分布，随后将其切片并打印在成像板上。对冠状动脉微循环最为重要的线粒体氧代谢和心肌血流分布，目前尚无先例研究一次性测量。在我们的研究中，我们还处理了高血压大鼠。通过NADH荧光研究，我们发现心肌微循环存在血流分布不均一性，缺血时血流分布不均一性增强。分子示踪法证实了这一点。两种方法的微循环最小单位长度大小相同，均为400微米左右。在高血压基础研究方面，与对照大鼠相比，不对称二甲基精氨酸（ADMA）水平与一氧化氮（NO）生成的关系显著升高。在糖尿病大鼠的NADH荧光研究中，从常氧状态到缺氧状态的时间常数有缩短的趋势，而从缺氧状态到常氧状态的时间常数有延长的趋势。这支持了糖尿病大鼠心脏易受缺血的影响。此外，缺血后再灌注血流与NO的产生密切相关。

项目成果

仲本博: "ラットにおける糖尿病冠循環とNO"Japanese Circulation Journal. 64・Suppl.. 647 (2000)

Hiroshi Nakamoto：“大鼠的糖尿病冠脉循环和 NO”日本循环杂志 64·Suppl.. 647 (2000)。

梶田達也: "NADH蛍光イメージングと分子トレーサイメージングを用いた局所心筋代謝と血流分布の空間的比較検討"電子情報通信学会技術報告. 99.177. 107-114 (1999)

Tatsuya Kajita：“利用 NADH 荧光成像和分子示踪成像进行局部心肌代谢和血流分布的空间比较研究” IEICE 99.177 (1999)。

矢田豊隆: "冠循環の統合的調節機構-高血圧と糖尿病における血管内障害の発症メカニズム-"医用電子と生体工学. 38. 14 (2000)

矢田丰贵：“冠脉循环的综合调节机制-高血压和糖尿病血管内疾病的发生机制”《医疗电子与生物工程》38. 14 (2000)。

Fumihiko Kajiya: "Intrarmyocardial influences on blood flow distributions in the myocardial wall"Annals of Biomedical Engineering. 28. 897-902 (2000)

Fumihiko Kajiya：“心肌内对心肌壁血流分布的影响”生物医学工程年鉴。

梶田達也: "ミトコンドリアNADH蛍光法による心筋の局所代謝機能評価-正常と糖尿病ラットでの比較"医用電子と生体工学. 37.suppl. 492 (1999)

Tatsuya Kajita：“通过线粒体 NADH 荧光法评估心肌的局部代谢功能 - 正常大鼠和糖尿病大鼠的比较”《医疗电子与生物工程》37.suppl.492（1999）。