Vanheel B, de Hemptinne A, Leusen I
Am J Physiol. 1986 May;250(5 Pt 1):C748-60. doi: 10.1152/ajpcell.1986.250.5.C748.
The influence of the surface pH (pHs) on the intracellular pH (pHi) and the recovery of pHi after an imposed intracellular acid load was investigated in isolated sheep cardiac Purkinje fiber, rabbit papillary muscle, and mouse and rat soleus muscle. pHs and pHi, respectively, were continuously measured by use of single- and double-barreled pH-sensitive glass microelectrodes. Surface acidosis, usually obtained by superfusion with solutions of acid pH, was also produced with low buffered (5 mM N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid) solutions at control pH. The pHs decrease (delta pHs) induced by low buffering was smallest (-0.08 pH unit) in Purkinje fiber and largest (-0.31 pH unit) in rat soleus muscle, which already had a more acid surface in control conditions. delta pHs was somewhat dependent on the superfusion rate. Higher superfusion rates decreased but did not abolish delta pHs. Surface acidosis was associated with a small intracellular acidification. Intracellular acid loads were produced by adding and subsequently withdrawing 20 meq/l NH4+ from the superfusate. In all preparations, the rate of recovery of pHi after NH4+ withdrawal was notably decreased at acidified pHs. This effect was amiloride sensitive. It is concluded that, in superfused multi-cellular preparations, pHs and therefore the buffer concentration of a superfusate can considerably influence steady-state pHi and pHi recovery from an imposed intracellular acid load.
在分离的绵羊心脏浦肯野纤维、兔乳头肌以及小鼠和大鼠比目鱼肌中,研究了表面pH值(pHs)对细胞内pH值(pHi)的影响以及施加细胞内酸负荷后pHi的恢复情况。分别使用单管和双管pH敏感玻璃微电极连续测量pHs和pHi。表面酸中毒通常通过用酸性pH值的溶液进行灌流来实现,也可在对照pH值下用低缓冲(5 mM N-2-羟乙基哌嗪-N'-2-乙烷磺酸)溶液产生。低缓冲诱导的pHs降低(ΔpHs)在浦肯野纤维中最小(-0.08个pH单位),在大鼠比目鱼肌中最大(-0.31个pH单位),而在对照条件下大鼠比目鱼肌的表面已经更酸。ΔpHs在一定程度上取决于灌流速率。较高的灌流速率会降低但不会消除ΔpHs。表面酸中毒与轻微的细胞内酸化有关。通过向灌流液中添加并随后去除20 meq/l的NH4+来产生细胞内酸负荷。在所有制剂中,酸化pH值时NH4+去除后pHi的恢复速率显著降低。这种效应是氨氯地平敏感的。得出的结论是,在灌流的多细胞制剂中,pHs以及因此灌流液的缓冲浓度可显著影响稳态pHi以及从施加的细胞内酸负荷中恢复的pHi。