非靶向代谢组学技术筛选成人大骨节病尿液差异代谢物

Screening of differential metabolites in urine of adult patients with Kashin-Beck disease by non-targeted metabonomics

目的筛选成人大骨节病患者尿液中差异代谢物及代谢通路,为寻找大骨节病特异性生物标志物及发病机制提供新的科学依据。方法在陕西省大骨节病病区永寿县选取成人大骨节病患者作为病例组,同时在同病区村选取无大骨节病临床症状的健康人群作为对照组。采集研究对象空腹中段晨尿,使用液相色谱-质谱联用(LC-MS)技术检测尿液中小分子代谢物,通过多变量统计分析偏最小二乘法判别分析(PLS-DA)以及与KEGG、人类代谢组学数据库(HMDB)比对,鉴定并筛选大骨节病患者差异代谢物及代谢通路。结果共纳入58名研究对象,病例组39名,其中男性23名、女性16名;年龄为(61.2±7.8)岁;体质指数为(22.7±6.5)kg/m^(2)。对照组19名,其中男性10名、女性9名;年龄为(50.0±9.0)岁;体质指数为(24.3±5.5)kg/m^(2)。鉴定并筛选出与大骨节病发病机制高度相关的一级差异代谢物3个(HT-2毒素、T-2四醇、硒-腺苷硒蛋氨酸),均表达下调;二级差异代谢物38个,其中10个上调、28个下调。筛选出差异代谢通路9条,主要涉及氨基酸代谢、脂类代谢和能量代谢。结论成人大骨节病患者和健康对照的尿液代谢谱存在显著差异,主要涉及氨基酸代谢、脂类代谢和能量代谢,其中一级差异代谢物HT-2毒素、T-2四醇、硒-腺苷硒蛋氨酸与大骨节病发病机制高度相关。

Objective To screen differential metabolites and metabolic pathways in urine of adult patients with Kashin-Beck disease(KBD),so as to provide scientific basis for finding specific biomarkers and pathogenesis of KBD.Methods In Yongshou County,the KBD area in Shaanxi Province,adult KBD patients were selected as the case group,and healthy people without clinical symptoms of KBD were selected as the control group in the same disease area.The subjects'fasting mid-morning urine was collected,and liquid chromatography-mass spectrometry(LC-MS)technology was used to detect small-molecule metabolites in the urine.Multivariate statistical analysis[partial least square discriminant analysis(PLS-DA)]and comparison with KEGG and human metabonomics database(HMDB)were used to identify and screen differential metabolites and metabolic pathways in KBD patients.Results A total of 58 subjects were included,39 cases in the case group,including 23 males and 16 females;the age was(61.2±7.8)years old;the body mass index was(22.7±6.5)kg/m^(2).There were 19 cases in the control group,including 10 males and 9 females;the age was(50.0±9.0)years old;the body mass index was(24.3±5.5)kg/m^(2).Three first-order differential metabolites(HT-2 toxin,T-2 tetraol and seleno-adenosine selenomethionine)were identified and screened,which were highly related to the pathogenesis of KBD,and all were down-regulated.There were 38 second-order differential metabolites,among them,10 were up-regulated and 28 were down-regulated.Nine differential metabolic pathways were screened,mainly involving amino acid metabolism,lipid metabolism and energy metabolism.Conclusions The urine metabolism profiles of adult KBD patients and healthy people are significantly different,mainly involving amino acid metabolism,lipid metabolism and energy metabolism.The first-order differential metabolites HT-2 toxin,T-2 tetraol and seleno-adenosine selenomethionine are highly correlated with the pathogenesis of KBD.