Analysis of Differential Metabolites in Selenium-enriched Rice Based on Extensive Non-targeted Metabolome

[Objectives]Selenium(Se)-enriched rice is the main type of Se-enriched agricultural product developed in China,and this study aimed to understand the impact of selenium application on the metabolites in rice.[Methods]Se-enriched rice was prepared by foliar application of a sodium selenite aqueous solution,and high-throughput analysis of differential metabolites in Se-enriched rice was conducted based on extensive non-targeted metabolome.[Results]There were significant differences in metabolites between Se-enriched rice and ordinary rice,and a total of 535 differential metabolites were identified.Among them,420 metabolites in Se-enriched rice were upregulated,accounting for 78.5%,far higher than downregulated metabolites.The enrichment differences of three KEGG metabolic pathways,including cysteine and methionine metabolism,zeatin biosynthesis,and arachidonic acid metabolism,reached a significant level,indicating that selenium enrichment had a significant regulatory effect on the metabolism of sulfur-containing amino acids,the synthesis of natural cytokinin zeatin,and arachidonic acid bioactive components in rice.[Conclusions]The results can provide a theoretical basis for the production of Se-enriched rice.

Baseline metabolites could predict responders with hepatitis B virus-related liver fibrosis for entecavir or combined with FuzhengHuayu tablet

BACKGROUND After receiving entecavir or combined with FuzhengHuayu tablet(FZHY)treatment,some sufferers with hepatitis B virus(HBV)-related liver fibrosis could achieve a histological improvement while the others may fail to improve even worsen.Serum metabolomics at baseline in these patients who were effective in treatment remain unclear.AIM To explore baseline serum metabolites characteristics in responders.METHODS A total of 132 patients with HBV-related liver fibrosis and 18 volunteers as healthy controls were recruited.First,all subjects were divided into training set and validation set.Second,the included patients were subdivided into entecavir responders(E-R),entecavir no-responders(E-N),FZHY+entecavir responders(FR),and FZHY+entecavir no-responders(F-N)following the pathological histological changes after 48 wk’treatments.Then,Serum samples of all subjects before treatment were tested by high performance liquid chromatographytandem mass spectrometry(LC-MS)high-performance LC-MS.Data processing was conducted using multivariate principal component analysis and orthogonal partial least squares discriminant analysis.Diagnostic tests of selected differential metabolites were used for Boruta analyses and logistic regression.RESULTS As for the intersection about differential metabolic pathways between the groups E-R vs E-N and F-R vs F-N,results showed that 4 pathways including linoleic acid metabolism,aminoacyl-tRNA biosynthesis,cyanoamino acid metabolism,alanine,aspartate and glutamate metabolism were screened out.As for the differential metabolites,these 7 intersected metabolites including hydroxypropionic acid,tyrosine,citric acid,taurochenodeoxycholic acid,benzoic acid,2-Furoic acid,and propionic acid were selected.CONCLUSION Our findings showed that 4 metabolic pathways and 7 differential metabolites had potential usefulness in clinical prediction of the response of entecavir or combined with FZHY on HBV fibrotic liver.

Identification of differential metabolites in liquid diet fermented with Bacillus subtilis using gas chromatography time of flight mass spectrometry

Growth and health responses of pigs fed fermented liquid diet are not always consistent and causes for this issue are still not very clear. Metabolites produced at different fermentation time points should be one of the most important contributors. However, currently no literatures about differential metabolites of fermented liquid diet are reported. The aim of this experiment was to explore the difference of metabolites in a fermented liquid diet between different fermentation time intervals. A total of eighteen samples that collected from Bacillus subtilis fermented liquid diet on days 7, 21 and 35 respectively were used for the identification of metabolites by gas chromatography time of flight mass spectrometry(GCTOF-MS). Fifteen differential metabolites including melibiose, sortitol, ribose, cellobiose, maltotriose,sorbose, iso maltose, maltose, fructose, D-glycerol-1-phosphate, 4-aminobutyric acid, beta-alanine, tyrosine, pyruvic acid and pantothenic acid were identified between 7-d samples and 21-d samples. The relative level of melibiose, ribose, maltotriose, D-glycerol-1-phosphate, tyrosine and pyruvic acid in samples collected on day 21 was significantly higher than that in samples collected on day 7(P < 0.01),respectively. Eight differential metabolites including ribose, sorbose, galactinol, cellobiose, pyruvic acid,galactonic acid, pantothenic acid and guanosine were found between 21-d samples and 35-d samples.Samples collected on day 35 had a higher relative level of ribose than that in samples collected on day 21(P < 0.01), In conclusion, many differential metabolites which have important effects on the growth and health of pigs are identified and findings contribute to explain the difference in feeding response of fermented liquid diet.

Metabolic profiles and morphological characteristics of leaf tips among different sweet potato(Ipomoea batatas Lam.)varieties

Sweet potato leaf tips have high nutritional value,and exploring the differences in the metabolic profiles of leaf tips among different sweet potato varieties can provide information to improve their qualities.In this study,a UPLC-Q-Exactive Orbitrap/MS-based untargeted metabolomics method was used to evaluate the metabolites in leaf tips of 32 sweet potato varieties.Three varieties with distinct overall metabolic profiles(A01,A02,and A03),two varieties with distinct profiles of phenolic acids(A20 and A18),and three varieties with distinct profiles of flavonoids(A05,A12,and A16)were identified.In addition,a total of 163 and 29 differentially expressed metabolites correlated with the color and leaf shape of sweet potato leaf tips,respectively,were identified through morphological characterization.Group comparison analysis of the phenotypic traits and a metabolite-phenotypic trait correlation analysis indicated that the color differences of sweet potato leaf tips were markedly associated with flavonoids.Also,the level of polyphenols was correlated with the leaf shape of sweet potato leaf tips,with lobed leaf types having higher levels of polyphenols than the entire leaf types.The findings on the metabolic profiles and differentially expressed metabolites associated with the morphology of sweet potato leaf tips can provide useful information for breeding sweet potato varieties with higher nutritional value.

Differential metabolite analysis of the pharmacodynamic differences between different ratios of Dahuang(Radix Et Rhizoma Rhei Palmati)-Taoren(Semen Persicae) herb pair

OBJECTIVE: To explore the material basis of the difference of efficacy of Dahuang(Radix Et Rhizoma Rhei Palmati)-Taoren(Semen Persicae)(DT) drugs with different proportions. METHODS: Samples of different ratios of Dahuang(Radix et Rhizoma Rhei Palnati, DH) to Taoren(Semen Persicae, TR)(Group A 1∶1, B 2∶3, C 3∶2) were analyzed based on gas chromatography time-of-flight mass spectrometry untargeted metabolomics technique. RESULTS: A total of 240 primary metabolites were detected. Forty-one differential metabolites involved nine differential metabolic pathways, of which four were closely related to the efficacy of DT in the treatment of heat and blood stasis syndrome. These pathways included the biosynthesis of amino acid(phenylalanine tyrosine and tryptophan), flavonoids, unsaturated fatty acids, and the glycolysis/glycogenesis pathway. CONCLUSION: There are significant differences in the efficacy of different ratios of DT drugs, and their optimal ratio for the treatment of heat and blood stasis syndrome should be 1∶1.

LC-MS-based metabolomics analysis of Gei Herba liver treatment in acetylphenylhydrazine-induced blood deficiency mice

Objective:This study aims to explore the metabolic mechanism of the liver protective effect of an aqueous extract of Gei Herba(AEG)in blood deficiency(BD)mice.Methods:The BD mouse model was established by acetylphenylhydrazine and cyclophosphamide.A total of forty-eight female Kunming mice(18–22 g)were randomly assigned to six groups:a control group,a BD model group,a positive drug group(Danggui Yixue oral liquid),and three AEG treatment groups receiving a daily AEG dose of 1,2,4 g/kg for nine days.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and the protein expression of IL-1,IL-2,IL-3 and TNF-αwere determined by enzyme-linked immunosorbent assay.The liquid chromatography-mass spectrometry(LC-MS)based metabolomics was used to identify liver metabolites,and the MetaboAnalyst platform was used to analyze the metabolic pathways.Results:AEG reduced serum AST,increased IL-1,IL-2,IL-3 and decreased TNF-αto effectively alleviate liver damage in BD mice.Furthermore,metabolomics analysis revealed that the disordered mice liver metabolic profile was corrected after AEG treatment,five differential metabolites were upregulated,and the mechanism was mainly related to increasing the primary bile acid biosynthesis.Conclusion:AEG has a regulatory effect on abnormal liver metabolism in BD mice.

Metabolomics analysis for skin ulceration syndrome of Apostichopus japonicus based on UPLC/Q-TOF MS

Skin ulceration syndrome(SUS)is the main diseases aff ected the development of sea cucumber(Apostichopus japonicus)culture industries.To better observe the changes in the sea cucumber A.japonicus with SUS and understand the pathogenesis of the disease,activities of superoxide dismutase(SOD),catalase(CAT),and level of malondialdehyde(MDA)in coelomic fluid were detected using the Assay Kit and metabolites in the body wall were assessed using ultra-performance liquid chromatography and quadrupole-time of flight mass spectrometry.The results indicated that level of MDA was increased during SUS compared with healthy individuals(P<0.01),but activities of SOD and CAT were reduced(P<0.05).In metabolomics analysis,metabolites,such as adenosine,choline,betaine aldehyde,palmitic acid,and taurine,were found to be upregulated and 2-oxoadipic acid,anthranilic acid(vitamin L1),thioetheramide-PC,cholesterol-3-sulfate,and pentadecanoic acid were downregulated(VIP>1 and P<0.1).Pathway enrichment analysis indicated most enrichment of KEGG pathways were mainly related to energy metabolism,immunity,and osmoregulation such as ABC transporters,glycine,serine and threonine metabolism,tryptophan metabolism and neuroactive ligand-receptor interaction.Our study reflected the difference in enzyme activity and metabolites between A.japonicus with SUS and those without,which will provide reference data for investigating SUS.

Study on metabonomics in patients with acute coronary syndrome based on LC-MS technology

Objective:To investigate the plasma metabolic components of patients with unstable angina(UA)and acute myocardial infarction(AMI),and to screen potential biomarkers and explore possible biological mechanisms in order to provide reference for early evaluation of acute coronary syndrome.Methods:Plasma samples from patients with UA and AMI were collected to obtain their general information,and the metabolites were detected by LCMS technology.Combined with univariate statistical analysis,the significantly different metabolites and their pathways were further determined by partial least squares discriminant analysis and orthogonalized partial least squares discriminant analysis models.Results:A total of 33 samples from UA group and 47 samples from AMI group were included for testing.Then 54 metabolites and 20 pathways were found to be significantly different between them.Metabolites such as pantoprazole,acetylcarnitine,palmitoyl ethanolamide,betaine,caprylic acid,bilirubin,histidine,oleic acid,citrate,vanillin can be used as potential biomarkers to distinguish them.The pathways include ABC transporters,aminoacyl-tRNA biosynthesis,central carbon metabolism in cancer and so on.Conclusion:There are significant differences in the plasma metabolic components of UA and AMI.Studies related to metabolomics may guide significance for exploring the biological nature of ACS.

Apocynum venetum Feed Affects the Serum Metabolism of Tan-sheep through the Amino Acid and Glycolipid Metabolic Pathways

[Objective]The paper was to explore the effects of Apocynum venetum diet on nutritional metabolism of Tan sheep.[Method]Forty Ningxia Tan sheep were randomly divided into 4 groups(A,B,C and D),with 10 sheep in each group.The sheep were fed with different contents of A.venetum granule feedstuff(0%,5%,10%and 15%).Blood samples were collected from jugular vein on the 20th,40th and 60th day of the experimental period.Serum samples were prepared and analyzed for differential metabolites by UPLC-Q-TOF/MS technique and annotated to the KEGG pathway.[Result](1)The differential metabolites increased with the extension of feeding time.The up-regulated metabolites in Tan sheep serum were more than the down-regulated ones,and reached the maximum on the 40th day.The up-regulated differential metabolites included 2 amino acids(valine and alanine),7 organic acids(β-hydroxybutyric acid,fumarate,ethylmalonic acid,hydroxyphenylacetic acid,gentiolic acid,protocatechuic acid,and oxycholic acid),pyrocatechol and taurochenocholate.The down-regulated differential metabolite was only p-chlorophenol.(2)With the increase of A.venetum content in pelleted diet,the differential metabolites in the serum of Tan sheep also increased,and the serum metabolic level gradually stabilized after the 10%concentration level.Similarly,the up-regulated metabolites were far more than the down-regulated metabolites.The up-regulated differential metabolites included 3 organic acids(β-hydroxybutyric acid,oxycholic acid and gentiolic acid),2 amino acids(alanine and valine)and catechol,and the down-regulated differential metabolite was only taurochenocholate.(3)The metabolic pathways involving in differential metabolites were mainly tricarboxylic acid cycle,tyrosine metabolism,taurine and bile acid pathway.[Conclusion]The results will provide a scientific basis for the green breeding of Tan sheep.