基于质谱技术的早产儿视网膜病血代谢产物的初步研究

Blood metabolites in preterm infants with retinopathy of prematurity based on tandem mass spectrometry: a preliminary study

摘要:

目的 应用液相色谱-串联质谱技术（liquid chromatography tandem mass spectrometry，LC-MS/MS）和代谢组学方法探讨早产儿视网膜病（retinopathy of prematurity，ROP）患儿出生血代谢产物差异，寻找ROP早期诊断的新的生物标记物。方法 收集2013年1月—2016年12月中山大学附属第六医院住院的21例ROP患儿（ROP组）及同期21例非ROP患儿（非ROP组）干血片标本，利用LC-MS/MS进行代谢产物测定，运用正交偏最小二乘判别分析法（orthogonal partial least squares discriminant analysis，OPLS-DA）寻找差异物质和生物标记物。结果 ROP组和非ROP组患儿血代谢谱有明显差异，经模式识别分析、代谢物得分图（Score-plot，S-plot）、权重分析初步得出10个差异较大的氨基酸。进一步统计分析发现ROP组患儿血谷氨酸、亮氨酸、天冬氨酸、鸟氨酸和甘氨酸水平明显高于非ROP组，差异有统计学意义（P<0.05）。受试者工作特征曲线分析显示，谷氨酸及鸟氨酸对ROP诊断价值最高。结论 ROP患儿与非ROP患儿比较血代谢产物具有明显差异，谷氨酸及鸟氨酸是诊断ROP的代谢标志物。LC-MS/MS结合代谢组学分析方法在ROP早期识别与诊断中具有潜在的应用价值。

Abstract:

Objective To study new biomarkers for the early diagnosis of retinopathy of prematurity (ROP) by analyzing the differences in blood metabolites based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and metabolomics.Methods Dried blood spots were collected from 21 infants with ROP (ROP group) and 21 infants without ROP (non-ROP group) who were hospitalized in the Sixth Affiliated Hospital of Sun Yat-sen University from January 2013 to December 2016. LC-MS/MS was used to measure the metabolites, and orthogonal partial least squares-discriminant analysis was used to search for differentially expressed metabolites and biomarkers.Results There was a significant difference in blood metabolic profiles between the ROP and non-ROP groups. The pattern recognition analysis, Score-plot, and weight analysis obtained 10 amino acids with a relatively large difference. Further statistical analysis showed that the ROP group had significant increases in blood levels of glutamic acid, leucine, aspartic acid, ornithine, and glycine compared with the non-ROP group (P<0.05). The receiver operating characteristic curve analysis showed that glutamic acid and ornithine had the highest value in diagnosing ROP.Conclusions Blood metabolites in preterm infants with ROP are different from those without ROP. Glutamic acid and ornithine are the metabolic markers for diagnosing ROP. LC-MS/MS combined with metabolomics analysis has a potential application value in the early identification and diagnosis of ROP.