摘要
目的探讨基于液质联用(LC/MS)的代谢组学技术在2型糖尿病肾病动物模型KK-Ay小鼠不同病程的诊断应用价值。方法试验组KK-Ay小鼠和对照组C57BL/6J小鼠各15只,从8周龄开始,每4周收集24 h尿液测尿微量白蛋白,乙醚麻醉后空腹采血,分离血清检测空腹血糖、甘油三酯、尿素氮、肌酐、白蛋白等,同时对血清样品预处理进行LC/MS检测及代谢组学分析。第8、12、20周龄两组各处死3只小鼠,灌洗取肾后进行病理学检查。结果12周龄后KK-Ay小鼠空腹血糖、甘油三酯、尿微量白蛋白与C57BL/6J小鼠有统计学差异(P<0.01)。KK-Ay小鼠12周龄系膜基质增加明显,20周龄出现基底膜增厚等病理改变。代谢组学主成份分析显示,各周龄KK-Ay小鼠与C57BL/6J小鼠血清代谢图谱差异显著,早于血生化和肾脏病理变化,代谢物模式的经时变化轨迹图能够反映2型糖尿病肾病的进展规律。结论基于LC/MS的代谢组学技术对动物血清的检测分析能准确区分各周龄试验组和对照组小鼠,并揭示病程进展规律,其作为一种辅助手段,可能在2型糖尿病肾病的早期诊断中具有应用前景。
Objective To access the capability of LC/MS based metabonomics in the evaluation of different course of diabetic nephropathy in KK-Ay mice. Methods Thirty male KK-Ay mice and C57BL/6J mice each were studied at the age of eight weeks. A 24-hour urine collection was made for the tests of protein urine every four weeks. Blood serum were collected for biochemical and LC/MS analysis. Three mice of each group were sacrificed at the age of 8, 12, 20 weeks. Renal tissue was observed to assess pathologic change. Results The serum glucose, lipids, urine protein were increased in KK-Ay mice after the age of 12 weeks（P 〈0.01 ）. There were significant mesangial matrix accumulation at the age of 12 weeks and GBM thickening at the age of 20 weeks. Principal component analysis （PCA） can clearly identify the difference between the serum samples of KK-Ay mice from the C57BL/6J mice. Changes of metabomomics were earlier than that in the pathology and biochemistry of serum, the footprint of the metabolites could indicate the course of diabetic nephropathy in KK-Ay mice. Conclusions LC/MS based metabonomics could distinguish the KK-Ay mice from the C57/BL6J accurately, and indicate the course of diabetic. It therefore may be a promising new technology in diagnosing the DN of T2DM earlier.
出处
《山东医药》
CAS
北大核心
2010年第9期19-22,共4页
Shandong Medical Journal
基金
上海市科委重大项目(08dz1900600)
关键词
糖尿病
2型
糖尿病肾病
代谢组学
液质联用
主成份分析
diabetes mellitus, type 2
diabetic nephropathy
Metabomomic
Liquid Chromatography-mass Spectrometry
principal component analysis
