Berthold H K, Crain P F, Gouni I, Reeds P J, Klein P D
Stable Isotope Laboratory, U.S. Department of Agriculture/Agricultural Research Service Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030, USA.
Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10123-7. doi: 10.1073/pnas.92.22.10123.
The absorption and metabolism of dietary nucleic acids have received less attention than those of other organic nutrients, largely because of methodological difficulties. We supplemented the rations of poultry and mice with the edible alga Spirulina platensis, which had been uniformly labeled with 13C by hydroponic culture in 13CO2. The rations were ingested by a hen for 4 wk and by four mice for 6 days; two mice were fed a normal diet and two were fed a nucleic acid-deficient diet. The animals were killed and nucleosides were isolated from hepatic RNA. The isotopic enrichment of all mass isotopomers of the nucleosides was analyzed by selected ion monitoring of the negative chemical ionization mass spectrum and the labeling pattern was deconvoluted by reference to the enrichment pattern of the tracer material. We found a distinct difference in the 13C enrichment pattern between pyrimidine and purine nucleosides; the isotopic enrichment of uniformly labeled [M + 9] isotopomers of pyrimidines exceeded that of purines [M + 10] by > 2 orders of magnitude in the avian nucleic acids and by 7- and 14-fold in the murine nucleic acids. The purines were more enriched in lower mass isotopomers, those less than [M + 3], than the pyrimidines. Our results suggest that large quantities of dietary pyrimidine nucleosides and almost no dietary purine nucleosides are incorporated into hepatic nucleic acids without hydrolytic removal of the ribose moiety. In addition, our results support a potential nutritional role for nucleosides and suggest that pyrimidines are conditionally essential organic nutrients.
与其他有机营养素相比,膳食核酸的吸收和代谢受到的关注较少,这主要是由于方法上的困难。我们用可食用藻类钝顶螺旋藻补充家禽和小鼠的日粮,该藻类通过在13CO2中进行水培而被13C均匀标记。母鸡摄入日粮4周,四只小鼠摄入6天;两只小鼠喂食正常日粮,两只喂食缺乏核酸的日粮。处死动物后,从肝脏RNA中分离出核苷。通过负离子化学电离质谱的选择离子监测分析核苷所有质量同位素异构体的同位素富集情况,并参照示踪物质的富集模式对标记模式进行反褶积分析。我们发现嘧啶核苷和嘌呤核苷的13C富集模式存在明显差异;在禽类核酸中,均匀标记的嘧啶核苷[M + 9]同位素异构体的同位素富集比嘌呤核苷[M + 10]高出2个数量级以上,在鼠类核酸中高出7倍和14倍。与嘧啶核苷相比,嘌呤核苷在质量低于[M + 3]的较低质量同位素异构体中富集程度更高。我们的结果表明,大量膳食嘧啶核苷且几乎没有膳食嘌呤核苷在不水解去除核糖部分的情况下被纳入肝脏核酸。此外,我们的结果支持核苷具有潜在的营养作用,并表明嘧啶是条件必需的有机营养素。