Plasma Metabonomics In Severe Pre-eclampsia

Preeclampsia (PE) is a common pregnancy-associated disorder that complicates8%of pregnancies. The disease is responsible for60-70,000maternal deaths worldwide per annum. The pathogenesis of severe preeclampsia is thought to involve gene, immunology, oxidative response and other factors. The Pathogenesis of preeclampsia is unclear, Now that the incidence of preeclampsia and origin of the Whole body oxidation-antioxidant imbalance, plasma inflammatory factors involved in endothelial injury,and further lead to systemic vascular endothelial dysfunction, vasoconstriction,end-organ, hypoperfusion, increased vascular permeability, which The clinical symptoms caused by preeclampsia.Metabonomics is a newly developed approach, as an integral part of the systems biology encompassing a number of omics sciences such as genomics, transcriptomics and proteomics, and has become one of the hottest subjects worldwide. Metabonomics uses multivariate statistical technique to analyze highly complex data sets generated by high-throughput spectroscopy such as mass spectrometry (MS) of biological samples to capture metabolic variations in response to genetic modifications and environmental stimuli and track the metabolic pathway. Metabolic products are the end products of gene expression. The products level is determined by the metabolic pathways and the role of the enzyme activity of enzymes. They are associated with the body’s physiology, pathology, and development states. Therefore, metabolomics provides a way to study diseases from other side.The technology of metabnomics is developing rapidly. Liquid chromatography-mass spectrometry is efficient, sensitive and accurate. It is widely used in the analysis of hard, polar compounds, thermally unstable compounds and macromolecular compounds (including proteins, peptides, polysaccharides, polymers, etc). It is one of the most promising metabolomics technologies.Liquid chromatography can reduce ion suppression effects. The products can be well separated by liquid chromatography through selecting appropriate chromatographic conditions according to the same physical and chemical properties of isomers. The appropriate method of liquid chromatography can get lower detection limits and reduce background noise. We could get higher quality mass spectrometry data by using appropriate method. There are few chromatography libraries which can be used currently. However, we can build our own database and analytical spectrum by using standard materials. The unknown metabolites can be identified relying on comparison with the standard materials.OBJECTIVETo investigate the metabolic changes in the human plasma with severe pre-eclampsia and the value of metabolomic technology in early detection and diagnosis of severe pre-eclampsia.The objective was to investigate the different metabolic profile associated with severe preeclampsia and normal pregnancy to explore the pathogenesis of preeclampsia and to study the metabolites biomarkers in the plasma of severe pre-eclampsia.METHODS1. The plasma samples thawed at room temperature, drawing100μl samples in the centrifuge tube, add300μl protein precipite with methanol, vortex mixing for1minute, using high-speed refrigerated centrifuge, at4℃Conditions, the12,000rpm centrifugation for15minutes, the supernatant filtered through water filter needle. After filtration, the samples were analyzed by HPLC-MS.2. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis:we separately set the condition of liquid chromatography, mass spectrometry conditions and operating modes. Samples were adding automatic by the machine. 3. Statistical methods:LC-MS raw data files were inputed into the Marker View TM software for data extraction and pre-treatment, using principal component analysis (PCA) method to observe the overall distribution of samples in each group. We use the principal component discriminant analysis (PCA-DA) to identify metabolic profiles contributed greatly to the metabolites between the groups. We can find the most relevant differences metabolites in severe preeclampsia by the principal component discriminant analysis. The data use T test which is in MarkerViewTM package.RESULTS51women with severe pre-eclampsia and45matched control were distinguished by metabolomic methods. And16biomarkers were identified. The decrease change of16potential biomarker peaks in severe pre-eclampsia plasma, contrast to the matched control,are peak89、95、98、118、135、157、187、191、215、240and the increase peaks are peak139、155、284、451、837、922.CONCLUSIONWe could analyze the metabolic profile changes in plasma of uncomplicated and severe preeclampsia pregnancies by HPLC-MS technology, indicating that the method is suitable for studying the pathogenesis of preeclampsia. It is distinguished between women with severe pre-eclampsia and with normal pregnant using established metabolomics methods. The biomarkers can better represent the metabonic characteristics of severe pre-eclampsia.These findings justify a prospective assessment of metabolomic technology as an early diagnosed tool for severe pre-eclampsia.OBJECTIVEBy analyzing the concentration of plasma free animo acid in uncomplicated pregnancy and severe preeclampsia, we explore the effect of these amino acids on the pathogenesis of severe preeclampsia and metablolic pathway.METHODS1. The groups of culture medium of placenta:uncomplicated pregnancy (n=10), and severe preeclampsia (n=12).2. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis:we separately set the condition of liquid chromatography, mass spectrometry conditions and operating modes. Samples were adding automatic by the machine.3. Statistical methods:LC-MS raw data files were imputed into the Analyst software for data extraction and pre-treatment and then the data was exported to EXCEL.The data was analyzed with T test and Mann-Whitney test by applying software SPSS13.0.RESULTS1. By HPLC-3200QTrap technology, the free amino acid profile changes in plasma of uncomplicated and severe preeclampsia pregnancies are detected.2. Compared with normal pregnancy women:the concentration of proline, ornithine, phenylalanine, valine, and arginine was significantly lower.CONCLUSIONOur research found the concentration of glutamate, glutamine, kynurenine and tryptophan had significantly changes in the uncomplicated pregnancies and severe preeclampsia.The study demonstrated that free amina acid plays an important role in the pathogenesis of severe preeclampsia. We could analyze the free amino acid profile changes in plasma of uncomplicated and severe preeclampsia pregnancies by HPLC-MS technology, indicating that the method is suitable for studying the pathogenesis of preeclampsia. These findings justify a prospective assessment of metabolomic technology as an early diagnosed tool for severe pre-eclampsia.1. We could analyze the metabolic profile changes in plasma of uncomplicated and severe preeclampsia pregnancies by HPLC-MS technology, indicating that the method is suitable for studying the pathogenesis of preeclampsia. 2. It is distinguished between women with severe pre-eclampsia and with normal pregnant using established metabolomics methods.3. The biomarkers can better represent the metabonic characteristics of severe pre-eclampsia.These findings justify a prospective assessment of metabolomic technology as an early diagnosed tool for severe pre-eclampsia.4. We could analyze the free amino acid profile changes in plasma of uncomplicated and severe preeclampsia pregnancies by HPLC-MS technology, indicating that the method is suitable for studying the pathogenesis of preeclampsia.