Research Of Tissue Metabolomics In Granulomatous Lobular Mastitis Based On HPLC/Q-TOF-MS

Objective:To compare and analyze the metabolites of granulomatous lobular mastitis and granuloma-adjacent tissues through the metabonomics platform and on the basis of high performance liquid chromatography/Quadrupole time-of-flight mass spectrometer(HPLC/Q-TOF-MS),explore the difference index of granulomatous lobular mastitis via screening differential metabolites and abnormal metabolic pathways that might be involved,and provide a new method for the treatment of granulomatous lobular mastitis.Method:28 tissue samples pathologically diagnosed as granulomatous lobular mastitis and 28 corresponding granuloma-adjacent tissue samples were collected in Thyroid and Breast Surgery Department,Hunan Provincial People’s Hospital from December 2019 to February 2021.The metabolites of inflamed tissue and granuloma-adjacent tissue samples were analyzed by HPLC/Q-TOF-MS.After preprocessing of the acquired original metabonomics data via Metaboscape 3.0 software such as noise elimination,peak extraction and normalization,the data were exported.Simca-P14.1 software was used to perform partial least squares discriminant analysis(PLS-DA),principal component analysis(PCA)and orthogonal partial least square discriminant analysis(OPLS-DA)for the data after preprocessing,and screen the differential metabolites between granulomatous mastitis tissues and granuloma-adjacent tissues in accordance with the standards of variable weight value(VIP>1),T-test(p<0.05)and fold change(FC>2).Metabo Analysis 4.0 software was used to analyze the metabolic pathway of differential metabolites,and find the metabolic pathway related to granulomatous lobular mastitis.Result:1.There was an obvious difference in metabolic profiling of the metabolites between granulomatous lobular mastitis tissues and granuloma-adjacent tissues.2.It was found from the acquired original metabonomics data that there was a significant difference in the metabolic profiling between granulomatous mastitis tissues(Group Y)and granuloma-adjacent tissues(Group N),and 21 potential differential metabolites were screened according to VIP>1,FC>2 and p<0.05.Among them,12 metabolites such as Lyso PC20:3,5Z,8Z,11Z＿,norverapamil hydrochloride,Lyso PC26:1＿u5Z＿,vitamin D2 3-glucuronic acid,nonyl alcohol-9,3-hydroxy-9-hexadecyl carnitine,α-tocopherol,aspartyl phenylalanine and sulfadiazine had an elevated expression level,while 9 metabolites such as 19-hydroxyandrost-4-ene-3,17-dione,3-alanine sulfite,N-acetyl-D-glucosamine,4-guanidinobutyric acid and hydroxybutyryl carnitine had a reduced expression level.3.The 5 metabolic pathways,including taurine and subtaurine metabolism,amino sugar and nucleotide sugar metabolism,steroid hormone biosynthesis,terpenoid skeleton biosynthesis,and cysteine and methionine metabolism,might be involved in the pathological process of granulomatous lobular mastitis.Conclusion:1.There is a remarkable difference in metabolic profiling of the metabolites between granulomatous lobular mastitis tissues and granuloma-adjacent tissues.2.There are 21 metabolites with a different expression level in granulomatous lobular mastitis tissues such as Lyso PC20:3,5Z,8Z,11Z＿,norverapamil hydrochloride,Lyso PC26:1＿u5Z＿,vitamin D2 3-glucuronic acid,19-hydroxyandrost-4-ene-3,17-dione,3-alanine sulfite,and N-acetyl-D-glucosamine;and 5 metabolic pathways such as taurine and subtaurine metabolism,amino sugar and nucleotide sugar metabolism,and steroid hormone biosynthesis are involved in the disease development process of granulomatous lobular mastitis.