基于非靶标代谢组学技术探讨慢性肾脏病3-4期湿浊证患者的特征

Characterization of Patients with Chronic Kidney Disease Stages 3 to 4(Syndrome of Dampness Turbidity)Based on Non-target Metabonomics

目的:基于非靶标代谢组学技术诊断慢性肾脏病(CKD)3-4期湿浊证患者尿液及血清的差异性标志物。方法:选取2019年6月1日至2019年12月31日上海中医药大学附属龙华医院肾病科收治的CKD 3~4期湿浊证患者60例作为观察组,健康体检者10例作为对照组,运用非靶标代谢组学方法进行观察研究。结果:代谢PCA得分值的空间分布表明观察组和对照组尿液、血清代谢产物,差异有统计学意义(P<0.05)。尿液正交偏最小二乘法-判别分析(OPLS-DA)置换检验结果提示:R^(2)Y=0.67>0,Q^(2)Y=0.43>0。17个差异代谢物排序后,经标准品鉴定出5个差异性生物标志物分别为:熊果苷、水杨苷、4氧代脯氨酸、胍基乙酸及肌酸,并且明确其变化方向和主要参与的代谢途径(糖酵解及精氨酸和脯氨酸代谢)。血清OPLS-DA置换检验结果提示:R^(2)Y=0.70>0,Q^(2)Y=0.50>0。16个差异代谢物排序后,经标准品鉴定差异性生物标志物分别为:丙酮酸、琥珀酸半醛、富马酸、柠檬酸、苹果酸和α珀酮戊二酸,并且明确其变化方向和代谢途径(三羧酸循环及丙氨酸、天冬氨酸和谷氨酸代谢)。结论:代谢组学方法能较好地提升CKD 3~4期湿浊证的诊断精确度,是CKD 3~4期湿浊证的良好诊断手段。

Objective:To identify the differential markers in the urine and serum of patients with chronic kidney disease stages 3 to 4(syndrome of dampness turbidity)based on non-target metabonomics.Methods:Sixty patients with chronic kidney disease stages 3 to 4(syndrome of dampness turbidity)treated in the Longhua Hospital affiliated to Shanghai University of Traditional Chinese Medicine from June 1 to December 31 in 2019 and 10 volunteers confirmed healthy by physical examination were selected as the observation group and the control group,respectively.Non-target metabonomics was employed to identify the differential markers.Results:The principal component analysis of metabolism scores showed that the metabolites in the urine and serum had significant differences between the observation group and the control group(P<0.05).The permutation test of the orthogonal partial least squares-discriminant analysis(OPLS-DA)of urine metabolites showed R^(2)Y=0.67>0 and Q^(2)Y=0.43>0.After ranking of the 17 differential metabolites,5 differential biomarkers were identified with the reference substances,including arbutin,salicin,4-oxoproline,guanidinoacetic acid,and creatine.Their transformation paths and main metabolic pathways(glycolysis and arginine and proline metabolism)were clarified.The permutation test of the OPLS-DA of serum metabolites showed R^(2)Y=0.70>0 and Q^(2)Y=0.50>0.After ranking of the 16 differential metabolites,the differential biomarkers were identified based on the reference substances,including pyruvic acid,succinic acid semialdehyde,fumaric acid,citric acid,malic acid,andα-ketoglutaric acid.Furthermore,their transformation paths and main metabolic pathways(tricarboxylic acid cycle and metabolism of alanine,aspartate,and glutamate)were predicted.Conclusion:Metabonomics can improve the diagnostic accuracy of chronic kidney disease stages 3 to 4(syndrome of dampness turbidity)and thus serves as a good diagnostic method for this disease.