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Establishment Of Ceramide And Dihydroceramide By Sequential Window Acquisition Of All Theoretical Fragment Ion Spectra High Resolution Mass Spectrometry And Its Application In Liver Fibrosis Of Alcoholic Liver Disease

Posted on:2022-11-22Degree:MasterType:Thesis
Country:ChinaCandidate:J W ShiFull Text:PDF
GTID:2504306758490174Subject:Digestive System Disease
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Background and Objective:The number of patients with liver cirrhosis is increasing,including viral cirrhosis,cholestatic cirrhosis,alcoholic cirrhosis,autoimmune cirrhosis,metabolic cirrhosis,drug-induced cirrhosis and primary cirrhosis.Alcoholic cirrhosis is one of the most common types of cirrhosis in clinic.In the early stage of liver disease,that is,liver fibrosis,the pathological process is reversible.Liver biopsy is the gold standard for the diagnosis of viral and alcoholic cirrhosis.Alcoholic liver disease involves significant metabolic changes,including endoplasmic reticulum stress,mitochondrial dysfunction,and inflammation.Insulin resistance and hormonal changes also contribute to the development of the disease.In recent years,disturbances in sphingolipid metabolism and ceramide accumulation have received increasing attention as etiologies of alcoholic and non-alcoholic liver disease,with the consequences of ethanol intake attributed in part to the role of sphingolipids.The rise of metabolomics has contributed to the understanding and elucidation of disease pathologies at the small molecule level.The application of targeted metabonomics to explore the changes of ceramide in the progression of liver fibrosis in alcoholic liver disease can lead to early detection and treatment of the disease.Therefore,the aim of this study was to establish a high-resolution mass spectrometry method for the detection of sphingolipid ceramides and dihydroceramides,and to explore the clinical value of different ceramides in the progression of liver fibrosis in alcoholic liver disease.Methods:A Method for the determination of ceramide and dihydroceramide by Sequential window acquisition of all theoretical fragment ion spectra-High resolution mass spectrometry(SWATH-HRMS)was established and validated in 311 subjects,including 82 healthy controls and 88 patients with alcoholic liver disease(35 patients with mild alcoholic liver disease in fibrosis stage F1-F2,53 patients with severe alcoholic liver disease in fibrosis stage F3-F4)and 141 patients of chronic hepatitis B fibrosis(52 patients of mild fibrosis in F1-F2 and89 patients of severe fibrosis in F3-F4).Between-group and within-group assessments were performed according to Mann-Whitney U tests and KruskalWallis K-W tests.Receiver operating characteristic curve(ROC)was used to analyze the relationship between 7 ceramides and 4 dihydroceramides with fibrosis in alcoholic liver disease.Spss26.0 and Graph Pad Prism version 9software for graphics rendering.P < 0.05 was considered statistically significant.Results:(1)Ceramide C22,C24 and dihydroceramide C24: 1 had significant differences among the healthy control group,mild alcoholic liver disease fibrosis group and severe alcoholic liver disease fibrosis group.Ceramide C22 and C24 were down-regulated and dihydroceramide C24: 1 was up-regulated with the progression of liver fibrosis in alcoholic liver disease.Ceramide C24 and dihydroceramide C24: 1 had good sensitivity and specificity in the diagnosis of mild fibrosis in ALD with an area under the ROC curve of 0.76 [95%(CI)0.66-0.85] and 0.75 [95%(CI)0.65-0.85],respectively.Ceramide C22,C24,and dihydroceramide C24: 1 had good sensitivity and specificity in the diagnosis of severe fibrosis in ALD with an area under the ROC curve of 0.95[95%(CI)0.91-0.99],0.97 [95%(CI)0.94-0.99],0.83[95%(CI)0.75-0.90]。(2)Ceramide C20,C22 and C24 were significantly different among the healthy control group,the mild fibrosis group and the severe fibrosis group,and ceramide C20,C22 and C24 were down-regulated with the progression of hepatitis B fibrosis.Ceramide C24 and C24: 1 had good sensitivity and specificity in the diagnosis of mild fibrosis in hepatitis B,and the area under the ROC curve was 0.81 [95%(CI)0.74-0.89] and 0.71 [95%(CI)0.62-0.80],respectively.Ceramide C20,C22 and C24 had good sensitivity and specificity in the diagnosis of severe fibrosis of hepatitis B,and the area under the ROC curve was 0.82 [95%(CI)0.75-0.88],0.88 [95%(CI)0.83-0.93] and 0.96[95percent(CI)0.93-0.99],respectively.Conclusion:(1)In this study,SWATH-HRMS was used to detect 7 kinds of ceramides and 4 kinds of dihydroceramides simultaneously,and was applied to the determination of liver fibrosis in alcoholic liver disease.(2)Ceramide C22 and C24 were down-regulated with the progression of liver fibrosis,dihydroceramide C24: 1 was up-regulated in the progression of liver fibrosis in alcoholic liver disease,and ceramide C20 was down-regulated with the progression of hepatitis B fibrosis.The above sphingolipids had good diagnostic efficacy.(3)Ceramide C22 and C24 are expected to be potential diagnostic markers of alcoholic liver disease and chronic hepatitis B liver fibrosis,and dihydroceramide C24: 1 is expected to be a potential specific diagnostic marker of alcoholic liver disease liver fibrosis.
Keywords/Search Tags:ALD, liver fibrosis, sphingolipids, ceramides, HRMS
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