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心筋のカルシウム過剰負荷と細胞障害との相関に関する研究

心肌钙超载与细胞损伤的相关性研究

基本信息

批准号：
62624511
负责人：
金出英夫
金额：
$ 0.96万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
1987
资助国家：
日本
起止时间：
1987 至无数据
项目状态：
已结题

项目摘要

〔I〕灌流ラット心臓の細胞障害と細胞質(Ca)iの関係を直接測定するために, Ca指示色素フラ2を用いる生体蛍光測定法を開発した.(1).Ca指示色素フラ2を取込ませる方法. ランゲンドルフ型灌流ラット心臓を, フラ2/AM(25μM)を含むCardioplegia灌流液を用いて, 37°C×60分間定圧灌流(1ml/min)し, 次いで, Krebs液で余分の色素を洗い出した.(2).心臓の心外膜表面蛍光測定は, 申請者の開発によるガラスファイバー法を用いた. すなわち同心円状ファイバーの内円を石英(径3.5mm)とし, 励起光340mmと380mmを交互に照射した. 外円をガラス(径7mm)とし, 510mmの蛍光を記録した. 340mm励起による510mm蛍光強度と, 380mm励起による510mm蛍光強度の比(340/380)を(Ca)iの指標とした.(3).フラ2を取込んだ心臓の特徴. (Ca)iが測定できる程度, すなわち340/380比情報を明らかに記録できる程度のフラ2を取込いだ灌流心臓の収縮力は, 色素を取り込まない対照心臓の収縮力の20%程度であった.〔II〕フラ2を取り込ませた心臓を用いてCaパラドックス実験を行った. 無Ca液灌流開始直後から340/380比は徐々に低下し, 1分後には定常状態となった. フラ2を取り込ませない場合には観察できた, 無Ca期の拘縮は, フラ2取り込み心臓では観察されなかった. 無Ca灌流5あるいは10分後にはCaを再灌流すると340/380比は急激に上昇し, 約1分で一定値を示した. Ca再灌流後もフラ2投与群では, 静止張力の上昇(拘縮)が認められず, フラ2非投与群が著じるしい上昇(拘縮)を見せることと好対照を示した. フラ2投与群と非投与群におけるCPKの灌流液への逸脱程度は, 同じであった. フラ2を取り込んだ心臓のCaパラドックス実験から, Ca再灌流に伴ってCa流入があり, (Ca)iは急上昇すること, しかし, いわゆる静止張力上昇(拘縮)と心筋障害には直接関係が少ないことが示唆された.

(I) Perfusion cardiomyocyte barrier cell assay (Ca) I direct the determination of the enzyme enzyme, Ca instructs the pigment to use the phosphorescence assay to start the test. (1) Ca indicates the pigment to determine the concentration of the enzyme. 2/AM (25 μ M) contains Cardioplegia perfusion solution, 37 °C × 60 min fixed perfusion (1ml/min), secondary temperature, Krebs solution residue and pigment washing. (2) The epicardial surface of the heart is measured by photoluminescence, and the applicant is required to use the method. The quartz (diameter 3.5mm) is in the center, and the light 340mm 380mm is interactively irradiated. The external equipment (path 7mm) is wide open, and the 510mm light record is very bad. 340mm excites the light intensity of 510mm, and 380mm activates the light intensity ratio of 510mm. (Ca) I indicates the intensity of light. (3). Please tell me that you have a special heart. (Ca) I measure the degree of stress. (Ca) I measure the strength of the heart. (II) I measure the strength of the heart, the color, the heart, the heart. Immediately after the start of Ca perfusion, the temperature of 340 percussion 380 was lower than that of normal, and after 1 minute, the steady state was abnormal. During the Ca period, you will find that your heart is in good condition, and your heart will be monitored. After 5 minutes of Ca perfusion, the number of patients with Ca reperfusion was higher than that of acute shock, and about 1 minute must indicate that it was higher than that of acute shock. After the Ca reperfused, there are two shots in the group, the force in the rest, the force in the group, the force in the group, and the pressure in the group. The degree of escape is the same as that of the perfusate of CPK, which is the same as that of the group. The Ca reperfusion was accompanied by the inflow of Ca into the heart, (Ca) I was in a hurry, and there was no stress in the heart. (Ca) I was in a hurry to stop the pain.