A study on mechanism related to enhanced glucose transport by elevated CaィイD12+ィエD1 concentration during contraction of skeletal muscle

骨骼肌收缩过程中 CaD12+D1 浓度升高与葡萄糖转运增强相关机制的研究

• 批准号: 09480017
• 负责人: TABATA Izumi
• 金额: $ 5.89万
• 依托单位: National Institute of Fitness and Sports (1999)National Institute of Health and Nutrition (1997-1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09480017/
• 关键词: muscle contraction; calcium; skeletal muscle; glucose metabolism; 糖取り込み速度; GLUT4

Firstly, we have developed a method for measuring cytosol CaィイD12+ィエD1 concentration in mammal skeletal muscle tissue. Changes in CaィイD12+ィエD1 concentration was estimated using double-beam fluorescent method with fura-2/AM as a fluorescent indicator which penetrates into cytosol of skeletal muscle (emittion: 340 nm and 380 nm, fluorescent 500 nm). Using a small and thin forelimb muscle named epitrochlearis from rat weighing 100-110 g, CaィイD12+ィエD1 concentration in the muscle was evidenced to be measured during long-term incubation and submaximal contraction. However, during tetani contractions, CaィイD12+ィエD1 concentration was not found to be measured due to artifact related to tenani.Since recent studies suggested that stimulated glucose transport activity by contraction is related to increased CaィイD12+ィエD1 concentration in skeletal muscle, we observed effects of physiological stimuli including insulin, submaximal contraction, and submaximal hypoxia that do not induce tetani, and pharma … More cological stimuli that affect CaィイD12+ィエD1 concentration in cell.We firstly observed elevation of CaィイD12+ィエD1 in the muscle during incubation with low concentration caffeine (3 mM), W-7(50μM), and submaximal hypoxia. As these stimuli are known to increase glucose transport activity in the muscle, we think that the method is valid and valuable to clarify mechanisms related to submaximal contraction-stimulated glucose transport activity in rat skeletal muscle.There was no change in CaィイD12+ィエD1 during incubation in AICAR known to stimulate AMP kinase that may intervene muscle-contraction induced increase in glucose transport activity. This result suggests that pathway related to CaィイD12+ィエD1 -enhanced glucose transport activity does not exist prior to AMP kinase.Insulin has been shown not to affect CaィイD12+ィエD1 level in the muscle until 20 min after initiation of incubation. However, we observed that CaィイD12+ィエD1 concentration did increase after 20 min incubation in insulin (2 mU/ml). This result may suggest that this method can also be used for the purpose of elucidating insulin-induced glucose transport activity in skeletal muscle. Less

首先，我们建立了一种测定哺乳动物骨骼肌组织胞浆Ca ~（2+）D_1浓度的方法。以Fura-2/AM为荧光指示剂，采用双光束荧光法测定骨骼肌细胞内Ca ~（2+）D_（12+）D_（1+）浓度的变化。Fura-2/AM的荧光波长为340 nm和380 nm，荧光波长为500 nm。使用来自体重100-110 g的大鼠的小而薄的前肢肌肉（称为上滑车肌），证明在长期孵育和次最大收缩期间测量肌肉中的Ca ~（2+）D_（12+）D_（1）浓度。然而，在强直收缩过程中，由于tenani相关的伪影，未发现Ca ~（2+）D_（12+）D_（12+）D_1浓度的测量。由于最近的研究表明，收缩刺激的葡萄糖转运活性与骨骼肌中Ca ~（2+）D_（12+）D_1浓度的增加有关，我们观察了生理刺激（包括胰岛素、次最大收缩和次最大缺氧）和药物对强直收缩的影响。 ...更多信息 本实验首次观察到低浓度咖啡因（3 mM）、W-7（50μM）和亚极量缺氧对骨骼肌细胞内Ca ~（2+）D_（12+）D_1浓度的影响。由于这些刺激是已知的，以增加葡萄糖转运活动的肌肉，我们认为该方法是有效的和有价值的，以澄清机制有关的次最大收缩刺激葡萄糖转运活动在大鼠skeletal muscle.There是没有变化的Ca ~（2+）D_1在孵育过程中AICAR已知刺激AMP激酶，可能会干预肌肉收缩诱导的葡萄糖转运活动的增加。这一结果表明，与Ca ~（2+）D_12 + Ca ~（2+）D_1增强葡萄糖转运活性相关的途径在AMP激酶之前并不存在，胰岛素已被证明在孵育开始后20 min才影响肌肉中Ca ~（2+）D_12 + Ca ~（2+）D_1水平。然而，我们观察到在胰岛素（2 mU/ml）中孵育20分钟后，Ca ~（2+）D12+ Ca ~（2+）D1浓度确实增加。这一结果可能表明，该方法也可用于阐明胰岛素诱导的骨骼肌葡萄糖转运活性。少

项目成果

Tabata I, Y Suzuki, T Fukunaga, Yokozeki, H Akima, K. Funato: "Resistance training affects GLUT-4 content in skeletal muscle of humans after 19days of head down bed rest"J. Appl. Physiol.. 86. 909-914 (1999)

Tabata I、Y Suzuki、T Fukunaga、Yokozeki、H Akima、K. Funato：“阻力训练会影响头低卧床休息 19 天后人类骨骼肌中 GLUT-4 的含量”J。

Tabata I,K Kawanaka,S Nagaoka S Sekiguchi. Y Ohira.: "Effects of space flight on GLUT-4 content in rat plantaris muscle" Int J Biometeorol. 41. 101-104 (1998)

Tabata I、K Kawanaka、S Nagaoka S Sekiguchi。

Kawanaka K,I Tabata,M Higuchi.: "More tetanic contractios are required for activating glucose transport maximally in trained muscle." Journal of Applied Physiology. 83. 429-433 (1997)

Kawanaka K,I Tabata,M Higuchi.：“需要更多的强直性收缩才能最大程度地激活受过训练的肌肉中的葡萄糖转运。”

Tabata I, Y Suzuki, T Fukunaga, T Yokozeki, H Akima, K Funato: "Resistance training affects GLUT-4 content in skeletal muscle of humans after 19 days of head down bed rest."Journal of Applied Physiology. 86. 909-914 (1999)

Tabata I、Y Suzuki、T Fukunaga、T Yokozeki、H Akima、K Funato：“阻力训练会影响低头卧床休息 19 天后人类骨骼肌中 GLUT-4 的含量。”《应用生理学杂志》。

Tabaa I,Y Suzuki,T Fukunaga,Yokozeki,H Akima,K Funato: "Resistance training affects GLUT-4 content in skeletal muscle of humansafter 19days of head down bed rest."Journal of Applied Physiology. 86. 909-914 (1999)

Tabaa I、Y Suzuki、T Fukunaga、Yokozeki、H Akima、K Funato：“低头卧床休息 19 天后，阻力训练会影响人类骨骼肌中的 GLUT-4 含量。”应用生理学杂志。