摘要
目的:应用9.4 T液态离体型高场强、高分辨率核磁共振频谱仪分析大鼠肝脏代谢物。方法:20只雄性Wistar大鼠牺牲后取出肝脏组织置于低温液氮冷藏保存。肝脏组织加入2 mmol·m L-1冰冷的高氯酸,经过研磨、匀浆、p H中和后移至低温高速离心机进行高速离心,除去高氯酸盐及肝脏匀浆后残渣,获取上清液置于真空环境冻干。冻干后的肝脏提纯物粉末,用精细天平称量20 mg,随机置于含有10 mmol·L-1或1的TMSP内标及重水的5 mm内径核磁共振测试管中,并使样本完全溶解后进行磁共振频谱检测。磁共振检测在离体型9.4 T高分辨率核磁共振频谱仪进行。提纯物频谱通过含有抑水脉冲的Bruker zgpr序列获得。频谱在频率域上经机器自带的XWINNMR(Bruker Gm BH)软件初步处理,后通过MestRe-c 4.3进一步分析。结果:含有合适内标及肝脏代谢物比例的测试管中,9.4 T离体型高分辨率核磁共振频谱能准确定位频谱零点,获得质量良好的高氯酸提纯大鼠肝组织氢质子频谱,并能准确探测并定位常规的肝脏各种水溶性代谢物。结论:9.4 T离体型高分辨率核磁共振频谱能用于分析肝脏各种不同的代谢物。
Objective： To study the liver metabolites using in vitro 9. 4T high resolution magnetic resonance spectroscopy.Methods： 20 male Wistar rats were sacrificed and the livers were dissected. Liver tissues were frenzied under the low temperature liquid nitrogen. Before spectrum inspection,all of liver tissues were mixed with 2 mmol·mL^- 1cold perchlorate acid. The mixtures were grinded,homogenate and neutralized to PH 7. 0. Then,the mixed liquors were centrifuged in high speed low temperature centrifuge for removing the perchlorate acid and residue after liver homogenate. The supernatant liquids were retained and lyophilized in a low temperature vacuum. Finally,the purified Liver powders were acquired. 20 mg extract powders were transferred into a 5 mm NMR test tube which contained internal standard and heavy water. The samples were made completely dissolved. Spectroscopy data were collected using in vitro 9. 4T high resolution nuclear magnetic resonance spectrometer. Spectra of extracts were acquired using Bruker zgpr pulse program which contained a water suppression pulse sequence. Spectroscopy data were preliminary processed in frequency domain by the machine＇s own XWINNMR（ Bruker Gm BH） software and further analyzed using MestRe-c 4. 3 software. Results： Spectroscopy zero was accurate positioning in contained suitable proportion of internal standard and liver metabolites test tubes. Good quality 1H proton spectroscopies of Liver powders purified perchloric acid were acquired. The various water-soluble metabolites of normal liver were detected and positioned. Conclusions： A variety of different metabolites liver can be detected by 9. 4T in vitro high resolution nuclear magnetic resonance spectroscopy. The liver spectroscopy is with low noise and has smooth baseline.
出处
《赣南医学院学报》
2015年第2期228-231,共4页
JOURNAL OF GANNAN MEDICAL UNIVERSITY
关键词
肝脏
9.4
T离体型核磁共振氢谱
代谢物
rat livers
9.4T in vitro nuclear magnetic resonance spectroscopy
metabolites
