Uchida K, Doi K
Department of Physiology, Yamagata University School of Medicine, Japan.
Respir Physiol. 1994 Jul;97(2):213-23. doi: 10.1016/0034-5687(94)90027-2.
ATP concentration ([ATP]) and the fraction of rod-shaped myocytes (Rrod) were measured for quiescent ventricular cells isolated from the rat heart. The myocytes were gently shaken with metabolic inhibitors for 30 min at 37 degrees C, and then [ATP] was assayed enzymatically. [ATP] (mean +/- SE in nmol.(10(6) cells)-1) of the control group (80.2 +/- 6.3) was significantly reduced to 32.3 +/- 3.6 and 21.9 +/- 4.5 by inhibiting glycolysis with 2-deoxyglucose (DG, 10 mM) and iodoacetic acid (IAA, 2 mM), respectively. Inhibition of the oxidative phosphorylation by sodium cyanide (CN, 2 mM) or 2,4-dinitrophenol (DNP, 40 microM) did not induce significant [ATP] reduction (56.5 +/- 5.6 or 53.2 +/- 7.5). Rrod were 54.4 +/- 1.9, 47.3 +/- 5.1, 43.3 +/- 3.3, 28.5 +/- 3.6 and 11.6 +/- 1.8% for the control, CN, DNP, DG and IAA groups, respectively. Under glycolytic inhibition with DG, addition of pyruvate (5 mM) with a pyruvate dehydrogenase activator (dichloroacetate (DCA, 1 mM) or (-)isoproterenol (ISO, 10 microM)) restored [ATP] to 52.5 +/- 8.3 and 67.7 +/- 7.2, respectively, but Rrod still remained depressed (34.6 +/- 2.1 and 35.4 +/- 1.3%). We conclude that the normal shape of quiescent myocytes is maintained primarily by glycolytic ATP.
对从大鼠心脏分离出的静息心室细胞测量了三磷酸腺苷(ATP)浓度([ATP])和杆状心肌细胞的比例(Rrod)。将心肌细胞与代谢抑制剂在37℃轻轻振荡30分钟,然后用酶法测定[ATP]。对照组的[ATP](以nmol·(10⁶个细胞)⁻¹为单位,平均值±标准误)为(80.2±6.3),分别用2-脱氧葡萄糖(DG,10 mM)和碘乙酸(IAA,2 mM)抑制糖酵解后,显著降低至32.3±3.6和21.9±4.5。用氰化钠(CN,2 mM)或2,4-二硝基苯酚(DNP,40 μM)抑制氧化磷酸化并未导致[ATP]显著降低(分别为56.5±5.6或53.2±7.5)。对照组、CN组、DNP组、DG组和IAA组的Rrod分别为54.4±1.9%、47.3±5.1%、43.3±3.3%、28.5±3.6%和11.6±1.8%。在DG抑制糖酵解的情况下,添加丙酮酸(5 mM)与丙酮酸脱氢酶激活剂(二氯乙酸(DCA,1 mM)或(-)异丙肾上腺素(ISO,10 μM))分别将[ATP]恢复至52.5±8.3和67.7±7.2,但Rrod仍保持降低状态(分别为34.6±2.1%和35.4±1.3%)。我们得出结论,静息心肌细胞的正常形态主要由糖酵解产生的ATP维持。