基于代谢组学的儿童大骨节病患者尿差异代谢物的筛选

Screening of different metabolites in urine of children with Kaschin-Beck disease by the metabonomics technique

目的 通过代谢组学方法筛选大骨节病（KBD）儿童尿液中的差异代谢物，为进一步研究KBD尿代谢生物标志物提供线索。方法 在青海省KBD病区7 - 15岁寄宿制学生中，通过临床检查和X线拍片诊断KBD，选择56例病区KBD儿童作为病例组，51例病区健康儿童作为内对照组；同时，在经济水平和生活习惯相似的非病区选择同年龄组健康儿童50例作为外对照组。采集观察对象随意尿样，应用超高效液相色谱-四级杆-飞行时间质谱仪检测尿样中相对分子质量小的代谢物，通过偏最小二乘法判别分析（PLS-DA）和多变量统计分析筛选出有意义的内源性差异代谢物。结果 3组间代谢物分离明显，病例组与内、外对照组比较，差异有统计学意义（P 〈 0.05），最终筛选出22种内源性差异代谢物。与外对照组比较，21种代谢物在病例组表现为上调，其中有14种代谢物（包括L-胱氨酸、异丁酰肉碱、丁酰氯化肉碱、天门冬氨酰-天冬酰胺、天冬酰胺-天门冬氨酸、辅酶Q-2等）在内对照组与病例组间比较，差异有统计学意义（P均 〈 0.05）；内、外对照组间比较，有4种代谢物差异有统计学意义（P均 〈 0.05）。结论 KBD儿童的尿代谢物与健康儿童不同，差异代谢物L-胱氨酸、异丁酰肉碱、丁酰氯化肉碱、天门冬氨酰-天冬酰胺、天冬酰胺-天门冬氨酸、辅酶Q-2与KBD关联较紧密，可为KBD生物标志物的筛选提供一定线索。

Objective To search the different metabolites related with Kaschin-Beck disease（KBD） in urine of children with KBD by metabolomics technique, in order to provide clues in researching KBD biomarkers. Methods Among the children aged 7 - 15 years old in the KBD areas in Qinghai Province, the urine samples of 56 KBD children （case group）, 51 health children from the same area as the control group （internal control group） and 50 health children from non diseased areas （external control group） were examined by quadrupole-time of flight mass spectrometry （UPLC/QTOF-MS）. Partial least squares discriminate analysis （PLS-DA） and multivariate statistical analysis were applied to explore and search the different endogenic metabolites associated with KBD. Results A obvious separation among three groups was observed, and the difference was significant among case group and internal, external control groups （P 〈 0.05）. Twenty-two different endogenic metabolites associated with KBD were found, compared with the external control group, 21 metabolites were increased in the case group（P 〈 0.05）. Among them, 14 kinds of metabolites （L-Cystine, isobutyryl carnitine, butyryl-L-carnitine, aspartyl-Asparagine, asparaginyl-Aspartate and ubiquinone Q2, etc.） showed significant differences between the case group and internal control group （P 〈 0.05）. There were 4 kinds of metabolites showing significant differences between the two control groups（P 〈 0.05）. Conclusion The metabolic profile in the urine of KBD children and healthy children is obviously different and the metabolites of L-Cystine, isobutyryl carnitine, butyryl-L-carnitine, aspartyl-Asparagine, asparaginyl-Aspartate and ubiquinone Q2 are more closely related with KBD, which can be considered as the clues in screening KBD biomarkers.