31P NMR spectroscopy, chemical analysis, and free Mg2+ of rabbit bladder and uterine smooth muscle.
31P NMR spectroscopy, chemical analysis, and free Mg2+ of rabbit bladder and uterine smooth muscle.
复制标题
DOI：
10.1016/s0021-9258(18)66886-3
复制
发表时间：
1986-11
期刊：
影响因子：
--
通讯作者：
M. Kushmerick;P. Dillon;R. Meyer;T. Brown;J M Krisanda;H. Sweeney
中科院分区：
文献类型：
--
作者：
M. Kushmerick;P. Dillon;R. Meyer;T. Brown;J M Krisanda;H. Sweeney
离体兔膀胱和子宫的 31P NMR 谱是在静息和刺激时在体外稳态动脉灌注下获得的。光谱包含七个主要峰：磷酸乙醇胺、sn-甘油(3)磷酸胆碱、无机磷酸盐 (Pi)、磷酸肌酸以及 ATP 的 γ、α 和 β 峰。对膀胱水提取物的化学分析、高压液相色谱和核磁共振波谱鉴定出许多其他成分，这些成分也对完整组织的光谱做出了贡献，但在完整组织的光谱中并未得到解析：UTP、GTP、UDP-Glc、NAD+、磷酸胆碱和 sn-甘油(3)磷酸乙醇胺。根据 Pi 峰的化学位移测量，未刺激的膀胱和子宫的细胞内 pH 值为 7.10 +/- 0.09 S.D。和 7.01 +/- 0.12 S.D.，分别。与横纹肌（猫二头肌和大鼠心肌）中观察到的化学位移相比，平滑肌中 β-ATP 峰的化学位移显着升高（-0.3 ppm）。在受刺激和缺血的膀胱中发现了 ADP 峰值。将灌注膀胱中观察到的核苷酸的化学位移校准为含有离子强度为 180 mM 的磷酸肌酸、Pi、ADP 和 ATP 的溶液中游离 Mg2+ 浓度的函数。我们对细胞内游离 Mg2+ 浓度得出以下估计值：子宫，0.40 mM；未刺激膀胱，0.46 mM；刺激和缺血膀胱，0.50 mM（来自 ATP 化学位移）和 0.45（来自 ADP 化学位移）；猫二头肌，1.5 mM；和大鼠心肌，1.4 mM。
31P NMR spectra of isolated rabbit bladder and uterus were obtained under steady-state arterial perfusion in vitro at rest and while stimulated. The spectra contained seven major peaks: phosphoethanolamine, sn-glycero(3)phosphocholine, inorganic phosphate (Pi), phosphocreatine, and the gamma, alpha, and beta peaks of ATP. Chemical analyses, high-pressure liquid chromatography, and NMR spectroscopy of aqueous extracts of bladders identified a number of other components that also made contributions to, but were not resolved in, the spectra of the intact tissues: UTP, GTP, UDP-Glc, NAD+, phosphocholine, and sn-glycero(3)phosphoethanolamine. Intracellular pH of unstimulated bladders and uteri, measured from the chemical shift of the Pi peak, was 7.10 +/- 0.09 S.D. and 7.01 +/- 0.12 S.D., respectively. The chemical shift of the beta-ATP peak in the smooth muscles was significantly upfield (-0.3 ppm) compared to the chemical shift observed in striated muscles (cat biceps and rat myocardium). An ADP peak was identified in stimulated and ischemic bladders. The chemical shifts of the nucleotides observed in perfused bladders were calibrated as a function of free Mg2+ concentration in solutions containing phosphocreatine, Pi, ADP, and ATP at an ionic strength of 180 mM. We derived the following estimates for the intracellular free Mg2+ concentration: uterus, 0.40 mM; unstimulated bladder, 0.46 mM; stimulated and ischemic bladder, 0.50 mM (from the ATP chemical shift) and 0.45 (from the ADP chemical shift); cat biceps, 1.5 mM; and rat myocardium, 1.4 mM.