Cox D W, Morris P G, Feeney J, Bachelard H S
Biochem J. 1983 May 15;212(2):365-70. doi: 10.1042/bj2120365.
A system has been developed for performing 31P-n.m.r. studies on cerebral tissues superfused in vitro, and gives results comparable with those reported from studies in vivo. Under optimal superfusion conditions [10 mM-glucose and O2/CO2 (19:1)] the tissue concentrations of phosphocreatine and ATP were calculated to be approx. 3.1 and 1.3 mumol/g respectively. When the glucose of the superfusing medium was lowered to 0.5 mM, slightly decreased sugar phosphate peaks were observed, but there was no detectable change in [ATP] or [phosphocreatine]. At 0.2 mM-glucose, significantly decreased concentrations of phosphocreatine and ATP were observed. Substitution of pyruvate plus malate for glucose did not decrease levels of phosphocreatine and ATP. When the superfusing medium was gassed with air/CO2 (19:1; 'mild hypoxia'), there was an appreciable fall in sugar phosphates and phosphocreatine with no detectable effect on ATP. In the presence of N2/CO2 (19:1; 'severe hypoxia', since O2 was not completely excluded), concentrations of phosphocreatine fell considerably, but with little effect on ATP. The results demonstrate the feasibility of studying cerebral energy metabolism in vitro using the non-invasive 31P-n.m.r. technique and are discussed in relation to the sensitivity of cerebral tissues to metabolic insults in vitro and in vivo.
已开发出一种系统，用于对体外灌注的脑组织进行31P核磁共振研究，其结果与体内研究报告的结果相当。在最佳灌注条件下[10 mM葡萄糖和O2/CO2（19:1）]，磷酸肌酸和ATP的组织浓度经计算分别约为3.1和1.3 μmol/g。当灌注培养基中的葡萄糖降至0.5 mM时，观察到磷酸糖峰略有下降，但[ATP]或[磷酸肌酸]没有可检测到的变化。在0.2 mM葡萄糖时，观察到磷酸肌酸和ATP的浓度显著下降。用丙酮酸加苹果酸替代葡萄糖并没有降低磷酸肌酸和ATP的水平。当用空气/CO2（19:1；“轻度缺氧”）对灌注培养基进行通气时，磷酸糖和磷酸肌酸明显下降，而对ATP没有可检测到的影响。在存在N2/CO2（19:1；“严重缺氧”，因为未完全排除O2）的情况下，磷酸肌酸的浓度大幅下降，但对ATP影响很小。结果证明了使用非侵入性31P核磁共振技术在体外研究脑能量代谢的可行性，并结合脑组织在体外和体内对代谢损伤的敏感性进行了讨论。
