Shiota M, Sugano T
Am J Physiol. 1986 Jul;251(1 Pt 1):C78-84. doi: 10.1152/ajpcell.1986.251.1.C78.
The isolated rat hindlimb was perfused with Krebs-bicarbonate buffer without erythrocytes and albumin in a flow-through mode at 32 degrees C, and the viability and metabolic characteristics of perfused skeletal muscle were examined. 1) With the flow rate at 15 ml X min-1 X leg-1, glucose and O2 uptake, lactate release, lactate-to-pyruvate ratio in effluent, and tissue creatine phosphate and adenine nucleotides remained constant at rest during perfusion for 90 min. The twitch tension changed little over perfusion. 2) When the leg was stimulated at a frequency below 0.5 Hz, the standard flow rate adequately delivered O2 to the perfused leg. Sciatic nerve stimulation enhanced glucose uptake in the absence of insulin. 3) The stimulatory effect of insulin on glucose uptake was observed with a concentration as low as 0.1 mU/ml, and maximal effect was at approximately mU/ml, with a nearly eightfold increase in glucose uptake. 4) Epinephrine and isoproterenol at a concentration of 0.5 nM stimulated lactate release, with maximal effect at 5 nM. The response to catecholamines was reversible and reproducible with a single preparation during the perfusion period of 120 min. The results indicated that the perfusion of hindlimb with a hemoglobin- and albumin-free medium is a convenient and reliable tool for the biochemical investigations of the integral function of hindlimb skeletal muscle.
将分离的大鼠后肢在32℃下以流通模式用不含红细胞和白蛋白的Krebs - 碳酸氢盐缓冲液灌注，并检测灌注骨骼肌的活力和代谢特征。1）流速为15 ml·min⁻¹·腿⁻¹时，在90分钟的灌注静息期内，葡萄糖和氧气摄取、乳酸释放、流出液中乳酸与丙酮酸的比值以及组织磷酸肌酸和腺嘌呤核苷酸保持恒定。抽搐张力在灌注过程中变化不大。2）当以低于0.5 Hz的频率刺激腿部时，标准流速能充分向灌注的腿部输送氧气。坐骨神经刺激在无胰岛素的情况下增强了葡萄糖摄取。3）观察到胰岛素对葡萄糖摄取的刺激作用，浓度低至0.1 mU/ml时即可出现，最大效应约在1 mU/ml，葡萄糖摄取增加近八倍。4）浓度为0.5 nM的肾上腺素和异丙肾上腺素刺激乳酸释放，5 nM时达到最大效应。在120分钟的灌注期内，对儿茶酚胺的反应在单一制剂中是可逆且可重复的。结果表明，用不含血红蛋白和白蛋白的培养基灌注后肢是用于后肢骨骼肌整体功能生化研究的一种方便且可靠的工具。
