Nakanishi T, Seguchi M, Tsuchiya T, Yasukouchi S, Takao A
Department of Pediatric Cardiology, Heart Institute of Japan, Tokyo Women's Medical College.
Circ Res. 1990 Jul;67(1):111-23. doi: 10.1161/01.res.67.1.111.
This study investigated developmental changes in the effect of acidosis on intracellular pH (pHi) and [Ca]i in the isolated heart and isolated myocyte preparations. The whole heart or myocytes of newborn (5-7 days old) and adult rabbits were loaded with the fluorescent pH indicator 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) or calcium indicator fura-2. Left ventricular pressure in the isolated heart preparation and the magnitude of cell contraction in the single-cell preparation were monitored. The heart and single cell were illuminated with excitation lights (340 and 380 nm, respectively, for fura-2 and 438 and 490 nm for BCECF). The intensity of fluorescence from the ventricular surface or from the cell was detected. [Ca]i was estimated from the following ratio: fluorescence at 505 nm during excitation at 340 nm/fluorescence at 505 nm during excitation at 380 nm. pHi was estimated from the following ratio: fluorescence at 530 nm during excitation at 490 nm/fluorescence at 530 nm during excitation at 438 nm. In the newborn, depression of contractile function during respiratory acidosis or metabolic acidosis was less than in the adult. Diastolic and systolic [Ca]i increased during respiratory acidosis in both the newborn and adult, and the net changes in [Ca]i were similar in the two age groups. During respiratory or metabolic acidosis, pHi decreased, but the decrease in the newborn was significantly less than in the adult. These data suggest that the greater resistance of the newborn myocardium to acidosis is due to the smaller change of pHi in this age group and not due to the difference in [Ca]i alteration.
本研究调查了酸中毒对离体心脏和离体心肌细胞制剂中细胞内pH值(pHi)和细胞内钙浓度([Ca]i)影响的发育变化。将新生(5 - 7日龄)和成年兔的全心或心肌细胞用荧光pH指示剂2',7'-双(2 - 羧乙基)-5(6)-羧基荧光素(BCECF)或钙指示剂fura - 2进行负载。监测离体心脏制剂中的左心室压力和单细胞制剂中细胞收缩的幅度。用激发光(分别为fura - 2的340和380 nm以及BCECF的438和490 nm)照射心脏和单细胞。检测来自心室表面或细胞的荧光强度。[Ca]i通过以下比率估算:340 nm激发时505 nm处的荧光/380 nm激发时505 nm处的荧光。pHi通过以下比率估算:490 nm激发时530 nm处的荧光/438 nm激发时530 nm处的荧光。在新生兔中,呼吸性酸中毒或代谢性酸中毒期间收缩功能的降低小于成年兔。在新生兔和成年兔中,呼吸性酸中毒期间舒张期和收缩期的[Ca]i均升高,且两个年龄组中[Ca]i的净变化相似。在呼吸性或代谢性酸中毒期间,pHi降低,但新生兔中的降低明显小于成年兔。这些数据表明,新生心肌对酸中毒的更大抵抗力是由于该年龄组中pHi的变化较小,而非由于[Ca]i改变的差异。