Plasma Metabonomic Changes In Adenomyosis

OBJECTIVE: To determine the plasma metabolic profile of patients with adenomyosis by ultra high performance liquid chromatograph coupled with Q Exactive quadrupole-Orbitrap high resolution mass spectrometer on the basis of non-target metabolomics to maximize the discovery of abnormally expressed metabolites.METHODS: Twenty-six patients with adenomyosis who underwent laparoscopic hysterectomy between December 2017 and June 2018 in DaLian Obstetrics and Gynecology Hospital were included in the experimental group.For all these patients,adenomyosis was confirmed by postoperative pathology and other diseases were excluded and no hormone treatment was given.26 age-matched healthy subjects with no adenomyosis as evidenced by color Doppler ultrasonography and magnetic resonance imaging were included in the control group.5ml morning fasting venous blood was collected and centrifuged at 1500 g and 4 °C for 10 mins.Supernatant was then collected and stored at-80°C.After pretreated with derivatization,shaking,centrifugation,and lyophilization,serum samples were subjected to metabolomics experiments by LC-MS based on non-target metabolomics.Peak area was extracted using the compound discovery software to determine the structure of the metabolites,and the names and peak areas were obtained for all detectable metabolites.Metabolomics data were used for t-test analysis,principal component analysis(PCA),and partial least squares discriminant analysis(PLS-DA)by MetaboAnalyst software to find the metabolites abnormally expressed in the blood of patients with adenomyosis.The Pathway Analysis module in MetaboAnalyst software was searched to find the metabolic pathways that are responsible for these abnormally expressed metabolites,and the pathogenesis of adenomyosis was explored through representative abnormal metabolic pathway(s).Logistic regression analysis of abnormally expressed metabolites was performed by SPSS25.0 software to find the optimal metabolomics diagnosis model for adenomyosis,and the logistic regression equation for the model was obtained.ROC curve was plotted for this model by SPSS software to find the best diagnostictarget with the highest sensitivity and specificity.RESULTS: 1?A total of 550 metabolites were detected in blood samples,of which119 were identified and quantified.A total of 13 metabolites,including alanine,palmitoylcarnitine,lysophosphatidylethanolamine(LPE),valerylcarnitine,serine,5-hydroxyindoleacetic acid,lysophosphatidylcholine(LPC),glutamine,hydroxyindoleacetic acid,were abnormally expressed with statistical significance(p <0.05),making them abnormally expressed metabolites.Metabolites including serine and threonine among others were downregulated and those including palmitoylcarnitine and lysophosphatidic acid(LPA)were upregulated in the experimental group.2 ? Thirteen abnormally expressed metabolites were searched and matched to 9metabolic pathways,of which 8 metabolic pathways were completely matched and 1was approximately matched.Among them,metabolic pathways of palmitoylcarnitine,aminoacyl-tRNA biosynthesis and LPA mostly contributed to the production of these abnormally expressed metabolites,and the metabolic pathways of palmitoylcarnitine,LPA,and serine-threonine were enhanced.3?A diagnosis method for adenomyosis with the highest sensitivity and specificity can be achieved by combining four abnormally expressed metabolits,i.e.alanine(X1),palmitoylcarnitine(X2),LPE(20:4)(X4),and LPE(18:0)(X10).A logistic regression equation for this four-metabolite diagnostic model is established.CONCLUSIONS:1.Plasma metabolic profile is abnormal in patients with adenomyosis and 13 metabolites are abnormally expressed,among which metabolites including serine and threonine are downregulated and metabolites including palmitoylcarnitine and LPA are upregulated in the experimental group.2.Abnormal metabolic pathways are found in patients with adenomyosis and a total of 9 abnormal metabolic pathways are retrieved.Among them,the metabolic pathways of palmitoylcarnitine,serine-threonine,and LPA are abnormally overexpressed.These abnormally expressed metabolic pathways may be related to the occurrence,development and progression of this disease.3.A diagnosis method for adenomyosis with the highest sensitivity and specificitycan be achieved by combining four abnormally expressed metabolits,i.e.alanine(X1),palmitoylcarnitine(X2),LPE(20:4)(X4),and LPE(18:0)(X10).The highest sensitivity and specificity(96.2% for both)of this model are achieved when variable R=0.55709.Area under the ROC curve is 0.997.This model contributes to the early diagnosis of adenomyosis.