通过高效液相色谱法用硝基苄基硫代肌苷证明肝脏和横膈膜中的腺苷储备。

Demonstration of the adenosine reservoirs with nitrobenzylthioinosine in liver and diaphragm by high-performance liquid chromatography.

作者信息

Jenkins R L, McDaniel H G, Grizzle W, Parrish S W, McDaniel H B

机构信息

Department of Medicine, University of Alabama, Birmingham 35294.

出版信息

J Chromatogr. 1988 Apr 29;426(2):249-58. doi: 10.1016/s0378-4347(00)81953-2.

DOI:10.1016/s0378-4347(00)81953-2

PMID:3392139

Abstract

Purine nucleotides, nucleosides, nucleobases, dinucleotides and nucleosides derivatives from acid-extracted rat liver and diaphragm were separated and quantitated by reversed-phase ion-pair high-performance liquid chromatography with a mobile phase composed of 90 mM potassium phosphate, 15 mM tetrabutylammonium hydroxide and a 1-30% methanol gradient. During 5 min of ischemia, adenine and guanine nucleotides decreased along with significant declines in NAD and increases in adenosine, inosine, hypoxanthine, xanthine, NADP and adenylosuccinate. Nitrobenzylthioinosine by gavage (5 mg/kg per day for five days) increased adenosine levels but without any alteration in nucleobase levels. Adenosine was shuttled to every available intracellular reservoir which included in declining order of magnitude GDP greater than adenosylhomocysteine greater than adenosine greater than ADP greater than AMP greater than IMP = XMP = GMP.

摘要

通过反相离子对高效液相色谱法，使用由90 mM磷酸钾、15 mM氢氧化四丁基铵和1-30%甲醇梯度组成的流动相，对酸提取的大鼠肝脏和膈肌中的嘌呤核苷酸、核苷、核碱基、二核苷酸和核苷衍生物进行分离和定量。在5分钟的缺血过程中，腺嘌呤和鸟嘌呤核苷酸减少，同时NAD显著下降，腺苷、肌苷、次黄嘌呤、黄嘌呤、NADP和腺苷酸琥珀酸增加。通过灌胃给予硝基苄硫基肌苷（每天5 mg/kg，持续五天）可提高腺苷水平，但核碱基水平无任何变化。腺苷被转运到每个可用的细胞内储存库，其数量级从大到小依次为GDP大于腺苷同型半胱氨酸大于腺苷大于ADP大于AMP大于IMP = XMP = GMP。