Takahashi E, Sato K, Endoh H, Xu Z L, Doi K
Department of Physiology, Yamagata University School of Medicine, Yamagata 990-9585, Japan.
Am J Physiol. 1998 Jul;275(1):H225-33. doi: 10.1152/ajpheart.1998.275.1.H225.
The purpose of the present study was to directly visualize radial gradients of intracellular PO2 in a single individual cardiomyocyte isolated from the rat ventricle. Microspectrophotometry with the use of cytosolic myoglobin as an oxygen probe was conducted at 410 nm. When the quiescent cell was incubated with 1 microM carbonyl cyanide m-chlorophenylhydrazone to increase oxygen consumption approximately eightfold, gradual decreases in myoglobin oxygen saturation (SMb) were demonstrated toward the core of the cell, whereas these decreases disappeared when the cell was treated with 2 mM NaCN. These results highlighted the importance of diffusional oxygen transport in determining intracellular oxygenation in cardiac cells. From the measured SMb, we assessed the profile of radial changes in intracellular PO2 at the mean SMb comparable to that in vivo ( approximately 0.5). Quite steep PO2 gradients were demonstrated in the vicinity of the sarcolemma that were rapidly attenuated toward the cell core. These radial profiles of intracellular PO2 demonstrate the significance of myoglobin-facilitated diffusion of oxygen. Furthermore, the shallow gradients of PO2 near the center of the cell might arise from partial depression of oxygen consumption near the cell core.
本研究的目的是直接观察从大鼠心室分离出的单个心肌细胞内细胞内PO2的径向梯度。使用细胞溶质肌红蛋白作为氧探针,在410nm处进行显微分光光度测定。当将静止细胞与1μM羰基氰化物间氯苯腙一起孵育以将耗氧量增加约八倍时,发现肌红蛋白氧饱和度(SMb)朝着细胞核心逐渐降低,而当细胞用2mM NaCN处理时,这些降低消失。这些结果突出了扩散性氧转运在决定心脏细胞内氧合作用中的重要性。根据测得的SMb,我们评估了与体内相当(约0.5)的平均SMb时细胞内PO2的径向变化情况。在肌膜附近显示出相当陡峭的PO2梯度,该梯度朝着细胞核心迅速衰减。这些细胞内PO2的径向分布情况证明了肌红蛋白促进氧扩散的重要性。此外,细胞中心附近PO2的浅梯度可能是由于细胞核心附近耗氧量的部分降低所致。