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活体豚鼠心肌糖原代谢的碳-13核磁共振研究

Carbon-13 nuclear magnetic resonance studies of myocardial glycogen metabolism in live guinea pigs.

作者信息

Neurohr K J, Gollin G, Neurohr J M, Rothman D L, Shulman R G

出版信息

Biochemistry. 1984 Oct 9;23(21):5029-35. doi: 10.1021/bi00316a031.

Abstract

Myocardial glycogen metabolism was studied in live guinea pigs by 13C NMR at 20.19 MHz. Open-chest surgery was used to expose the heart, which was then positioned within a solenoidal radio frequency coil for NMR measurements. The time course of myocardial glycogen synthesis during 1-h infusions of 0.5 g of D-[1-13C]glucose (and insulin) into the jugular vein was investigated. The possible turnover of the 13C-labeled glycogen was also studied in vivo by following the labeled glucose infusion with a similar infusion of unlabeled glucose. The degree of 13C enrichment of the C-1 glycogen carbons during these infusions was measured in heart extracts by 1H NMR at 360 MHz. High-quality proton-decoupled 13C NMR spectra of the labeled C-1 carbons of myocardial glycogen in vivo were obtained in 1 min of data accumulation. This time resolution allowed measurement of the time course of glycogenolysis of the 13C-labeled glycogen during anoxia by 13C NMR in vivo. With the solenoidal coil used for 13C NMR, the spin-lattice relaxation time of the labeled C-1 carbons of myocardial glycogen could be measured in vivo. For a comparison, spin-lattice relaxation times of heart glycogen were measured in vitro at 90.55 MHz. Natural abundance 13C NMR studies of the quantitative hydrolysis of extracted heart glycogen in vitro at 90.55 MHz showed that virtually all the carbons in heart glycogen contribute to the 13C NMR signals. The same result was obtained in 13C NMR studies of glycogen hydrolysis in excised guinea pig heart.

摘要

采用20.19 MHz的¹³C核磁共振技术,在活体豚鼠身上研究了心肌糖原代谢。通过开胸手术暴露心脏,然后将其置于螺线管射频线圈内进行核磁共振测量。研究了经颈静脉输注0.5 g D-[1-¹³C]葡萄糖(和胰岛素)1小时期间心肌糖原合成的时间进程。还通过在输注标记葡萄糖后再输注类似剂量的未标记葡萄糖,在体内研究了¹³C标记糖原的可能周转情况。在这些输注过程中,通过360 MHz的¹H核磁共振测量心脏提取物中糖原C-1碳的¹³C富集程度。在1分钟的数据采集时间内,获得了体内心肌糖原标记C-1碳的高质量质子去耦¹³C核磁共振谱。这种时间分辨率使得能够通过体内¹³C核磁共振测量缺氧期间¹³C标记糖原的糖原分解时间进程。使用用于¹³C核磁共振的螺线管线圈,可以在体内测量心肌糖原标记C-1碳的自旋晶格弛豫时间。作为比较,在体外90.55 MHz下测量了心脏糖原的自旋晶格弛豫时间。在90.55 MHz下对提取的心脏糖原进行体外定量水解的天然丰度¹³C核磁共振研究表明,心脏糖原中几乎所有的碳都对¹³C核磁共振信号有贡献。在切除的豚鼠心脏中进行的糖原水解¹³C核磁共振研究也得到了相同的结果。

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