Objective:As an important part of metabolomics analysis,untargeted metabolomics has become a powerful tool in the study of tumor mechanisms and the discovery of metabolic markers with high-throughput spectrometric dat...Objective:As an important part of metabolomics analysis,untargeted metabolomics has become a powerful tool in the study of tumor mechanisms and the discovery of metabolic markers with high-throughput spectrometric data which also poses great challenges to data analysis,from the extraction of raw data to the identification of differential metabolites.To date,a large number of analytical tools and processes have been developed and constructed to serve untargeted metabolomics research.The different selection of analytical tools and parameter settings lead to varied results of untargeted metabolomics data.Our goal is to establish an easily operated platform and obtain a repeatable analysis result.Methods:We used the R language basic environment to construct the preprocessing system of the original data and the LAMP(Linux+Apache+MySQL+PHP)architecture to build a cloud mass spectrum data analysis system.Results:An open-source analysis software for untargeted metabolomics data(openNAU)was constructed.It includes the extraction of raw mass data and quality control for the identification of differential metabolic ion peaks.A reference metabolomics database based on public databases was also constructed.Conclusions:A complete analysis system platform for untargeted metabolomics was established.This platform provides a complete template interface for the addition and updating of the analysis process,so we can finish complex analyses of untargeted metabolomics with simple human-computer interactions.The source code can be downloaded from https://github.com/zjuRong/openNAU.展开更多
BACKGROUND In recent years,the incidence of type 2 diabetes(T2DM)has shown a rapid growth trend.Goto Kakizaki(GK)rats are a valuable model for the study of T2DM and share common glucose metabolism features with human ...BACKGROUND In recent years,the incidence of type 2 diabetes(T2DM)has shown a rapid growth trend.Goto Kakizaki(GK)rats are a valuable model for the study of T2DM and share common glucose metabolism features with human T2DM patients.A series of studies have indicated that T2DM is associated with the gut microbiota composition and gut metabolites.We aimed to systematically characterize the faecal gut microbes and metabolites of GK rats and analyse the relationship between glucose and insulin resistance.AIM To evaluate the gut microbial and metabolite alterations in GK rat faeces based on metagenomics and untargeted metabolomics.METHODS Ten GK rats(model group)and Wistar rats(control group)were observed for 10 wk,and various glucose-related indexes,mainly including weight,fasting blood glucose(FBG)and insulin levels,homeostasis model assessment of insulin resistance(HOMA-IR)and homeostasis model assessment ofβcell(HOMA-β)were assessed.The faecal gut microbiota was sequenced by metagenomics,and faecal metabolites were analysed by untargeted metabolomics.Multiple metabolic pathways were evaluated based on the differential metabolites identified,and the correlations between blood glucose and the gut microbiota and metabolites were analysed.RESULTS The model group displayed significant differences in weight,FBG and insulin levels,HOMA-IR and HOMA-βindexes(P<0.05,P<0.01)and a shift in the gut microbiota structure compared with the control group.The results demonstrated significantly decreased abundances of Prevotella sp.CAG:604 and Lactobacillus murinus(P<0.05)and a significantly increased abundance of Allobaculum stercoricanis(P<0.01)in the model group.A correlation analysis indicated that FBG and HOMA-IR were positively correlated with Allobaculum stercoricanis and negatively correlated with Lactobacillus murinus.An orthogonal partial least squares discriminant analysis suggested that the faecal metabolic profiles differed between the model and control groups.Fourteen potential metabolic biomarkers,including glycochenodeoxycholic acid,uric acid,13(S)-hydroxyoctadecadienoic acid(HODE),N-acetylaspartate,β-sitostenone,sphinganine,4-pyridoxic acid,and linoleic acid,were identified.Moreover,FBG and HOMA-IR were found to be positively correlated with glutathione,13(S)-HODE,uric acid,4-pyridoxic acid and allantoic acid and negatively correlated with 3-α,7-α,chenodeoxycholic acid glycine conjugate and 26-trihydroxy-5-β-cholestane(P<0.05,P<0.01).Allobaculum stercoricanis was positively correlated with linoleic acid and sphinganine(P<0.01),and 2-methyl-3-hydroxy-5-formylpyridine-4-carboxylate was negatively associated with Prevotella sp.CAG:604(P<0.01).The metabolic pathways showing the largest differences were arginine biosynthesis;primary bile acid biosynthesis;purine metabolism;linoleic acid metabolism;alanine,aspartate and glutamate metabolism;and nitrogen metabolism.CONCLUSION Metagenomics and untargeted metabolomics indicated that disordered compositions of gut microbes and metabolites may be common defects in GK rats.展开更多
BACKGROUND Nonalcoholic fatty liver disease(NAFLD)has become one of the most common chronic liver diseases in the world.In our early clinical data and questionnaire analysis of NAFLD,it was found that the body mass in...BACKGROUND Nonalcoholic fatty liver disease(NAFLD)has become one of the most common chronic liver diseases in the world.In our early clinical data and questionnaire analysis of NAFLD,it was found that the body mass index of some patients did not meet the diagnostic criteria for overweight or obesity.The consumption of high-temperature-processed foods such as fried food,hot pot and barbecue is closely related to the occurrence of nonobese NAFLD.Reducing the intake of this kind of food can reduce disease severity and improve prognosis.AIM To explore the untargeted metabolomics characteristics of nonobese nonalcoholic fatty liver disease in Sprague-Dawley rats induced by high-temperatureprocessed feed.METHODS Fifty-four male Sprague-Dawley rats were divided into three groups:The control group received a standard diet;the nonfried soybeans(NDFS)group received 60%NDFS and 40%basic feed and the dry-fried soybeans(DFS)group received 60%DFS and 40%basic feed.Six rats were sacrificed at week 4,8,and 12 in each group.The food intake,body weight,Lee’s index,liver index,serological index and hepatic histopathology were assessed.Untargeted metabolomics characteristics were used to analyze the changes in liver metabolites of rats at week 12.Correlations between metabolites and pathology scores between the DFS and control groups and between the DFS and NDFS groups were analyzed.We selected some of the metabolites,both within the pathway and outside of the pathway,to explain preliminarily the difference in liver pathology in the three groups of rats.RESULTS There were no statistically significant differences in the food intake,body weight,Lee's index or serological index between the DFS group and the control group(P>0.05).At week 8 and week 12,the steatosis scores in the DFS group were significantly higher than those in the other two groups(P<0.05).At week 12,the liver index of the DFS group was the lowest(NDFS group vs DFS group,P<0.05).The fibrosis score in the DFS group was significantly higher than those in the other two groups(P<0.05).The correlation analysis of the liver pathology score and differential metabolites in the DFS and NDFS groups showed that there were 10 strongly correlated substances:Five positively correlated substances and five negatively correlated substances.The positively correlated substances included taurochenodeoxycholate-3-sulfate,acetylcarnitine,20a,22bdihydroxycholesterol,13E-tetranor-16-carboxy-LTE4 and taurocholic acid.The negatively correlated substances included choline,cholesterane-3,7,12,25-tetrol-3-glucuronide,nicotinamide adenine dinucleotide phosphate,lysoPC[16:1(9Z)]and glycerol 3-phosphate.The correlation analysis of the liver pathology score and differential metabolites in the DFS and control groups showed that there were 13 strongly correlated substances:Four positively correlated substances and 9 negatively correlated substances.The positively correlated substances included 4-hydroxy-6-eicosanone,3-phosphoglyceric acid,13-hydroxy-9-methoxy-10-oxo-11-octadecenoic acid and taurochenodeoxycholate-3-sulfate.The negatively correlated substances included lysoPC[16:1(9Z)],S-(9-hydroxy-PGA1)-glutathione,lysoPC[20:5(5Z,8Z,11Z,14Z,17Z)],SM(d18:1/14:0),nicotinamide adenine dinucleotide phosphate,5,10-methylene-THF,folinic acid,N-lactoylglycine and 6-hydroxy-5-methoxyindole glucuronide.CONCLUSION We successfully induced liver damage in rats by using a specially prepared hightemperature-processed feed and explored the untargeted metabolomics characteristics.展开更多
Quality control of ginseng currently is mainly based on ginsenoside analysis,but rarely focuses on the volatile organic components.In the current work,an untargeted metabolomics approach,by headspace solid-phase micro...Quality control of ginseng currently is mainly based on ginsenoside analysis,but rarely focuses on the volatile organic components.In the current work,an untargeted metabolomics approach,by headspace solid-phase micro-extraction gas chromatography/mass spectrometry(HS-SPME-GC/MS),was elaborated and further employed to holistically compare the compositional difference of the volatile components simultaneously from 12 Panax herbal medicines,which included P.ginseng(PG),P.quinquefolius(PQ),P.notoginseng(PN),red ginseng(PGR),P.ginseng leaf(PGL),P.quinquefolius leaf(PQL),P.notoginseng leaf(PNL),P.ginseng flower(PGF),P.quinquefolius flower(PQF),P.notoginseng flower(PNF),P.japonicus(PJ),and P.japonicus var.major(PJvm).Chromatographic separation was performed on an HP-5MS elastic quartz capillary column using helium as the carrier gas,enabling good resolution within 1 h.We were able to characterize totally 259 volatile compounds,including 82 terpenes(T),46 alcohols(Alc),29 ketones(K),25 aldehydes(Ald),21 esters(E),and the others.By analyzing 90 batches of ginseng samples based on the untargeted metabolomics workflows,236 differential ions were unveiled,and accordingly 36 differential volatile components were discovered.It is the first report that simultaneously compares the compositional difference of volatile components among 12 Panax herbal medicines,and useful information is provided for the quality control of ginseng aside from the well-known ginsenosides.展开更多
Polymerase-tautomeric model for untargeted delayed base substitution mutations is proposed.Structural analysis of bases insertion showed that any canonical bases may be inserted opposite rare tautomeric forms of thymi...Polymerase-tautomeric model for untargeted delayed base substitution mutations is proposed.Structural analysis of bases insertion showed that any canonical bases may be inserted opposite rare tautomeric forms of thymine T3*,adenines A2*and A4*so that between them hydrogen bonds are formed.Canonical adenine and cytosine can be incorporated opposite canonical thymine only.Canonical thymine and guanine can be incorporated opposite canonical adenine only.If in the synthesis of DNA containing rare tautomeric forms of thymine T3*,adenines A2*and A4*,involved DNA polymerases with relatively high fidelity of synthesis,mutations not appear.However,if further DNA synthesis will involve DNA polymerases having a low fidelity of synthesis,there may be base substitution mutations.It was shown that the conclusion made in the Tomasetti and Vogelstein cancer risk model that the formation of about 67%of all mutations was not caused by exposure to any mutagens is erroneous.展开更多
BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therap...BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therapeutic effects and the possible mechanism of JPGS on DN.METHODS We first evaluated the therapeutic potential of JPGS on a DN mouse model.We then investigated the effect of JPGS on the renal metabolite levels of DN mice using non-targeted metabolomics.Furthermore,we examined the effects of JPGS on c-Jun N-terminal kinase(JNK)/P38-mediated apoptosis and the inflammatory responses mediated by toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3).RESULTS The ameliorative effects of JPGS on DN mice included the alleviation of renal injury and the control of inflammation and oxidative stress.Untargeted metabolomic analysis revealed that JPGS altered the metabolites of the kidneys in DN mice.A total of 51 differential metabolites were screened.Pathway analysis results indicated that nine pathways significantly changed between the control and model groups,while six pathways significantly altered between the model and JPGS groups.Pathways related to cysteine and methionine metabolism;alanine,tryptophan metabolism;aspartate and glutamate metabolism;and riboflavin metabolism were identified as the key pathways through which JPGS affects DN.Further experimental validation showed that JPGS treatment reduced the expression of TLR4/NF-κB/NLRP3 pathways and JNK/P38 pathway-mediated apoptosis related factors.CONCLUSION JPGS could markedly treat mice with streptozotocin(STZ)-induced DN,which is possibly related to the regulation of several metabolic pathways found in kidneys.Furthermore,JPGS could improve kidney inflammatory responses and ameliorate kidney injuries in DN mice via the TLR4/NF-κB/NLRP3 pathway and inhibit JNK/P38 pathwaymediated apoptosis in DN mice.展开更多
In the present study, total membrane proteins from tumor cell lines including HepG2, Hep3 B2,H226, Ovcar3 and N87 were extracted and digested with γLys C and trypsin. The resulting peptide lysate were pre-fractionate...In the present study, total membrane proteins from tumor cell lines including HepG2, Hep3 B2,H226, Ovcar3 and N87 were extracted and digested with γLys C and trypsin. The resulting peptide lysate were pre-fractionated and subjected to untargeted quantitative proteomics analysis using a high resolution mass spectrometer. The mass spectra were processed by the Max Quant and the protein abundances were estimated using total peak area(TPA) method. A total of 6037 proteins were identified, and the analysis resulted in the identification of 2647 membrane proteins. Of those, tumor antigens and absorption,metabolism, disposition and elimination(ADME) proteins including UDP-glucuronosyltransferase,cytochrome P450, solute carriers and ATP-binding cassette transporters were detected and disclosed significant variations among the cell lines. The principal component analysis was performed for the cluster of cell lines. The results demonstrated that H226 is closely related with N87, while Hep3 B2 aligned with HepG2. The protein cluster of Ovcar3 was apart from that of other cell lines investigated. By providing for the first time quantitative untargeted proteomics analysis, the results delineated the expression profiles of membrane proteins. These findings provided a useful resource for selecting targets of choice for anticancer therapy through advancing data obtained from preclinical tumor cell line models to clinical outcomes.展开更多
Abdominal aortic aneurysm(AAA)and atherosclerosis(AS)have considerable similarities in clinical risk factors and molecular pathogenesis.The aim of our study was to investigate the differences between AAA and AS from t...Abdominal aortic aneurysm(AAA)and atherosclerosis(AS)have considerable similarities in clinical risk factors and molecular pathogenesis.The aim of our study was to investigate the differences between AAA and AS from the perspective of metabolomics,and to explore the potential mechanisms of differential metabolites via integration analysis with transcriptomics.Plasma samples from 32 AAA and 32 AS patients were applied to characterize the metabolite profiles using untargeted liquid chromatography-mass spectrometry(LC-MS).A total of 18 remarkably different metabolites were identified,and a combination of seven metabolites could potentially serve as a biomarker to distinguish AAA and AS,with an area under the curve(AUC)of0.93.Subsequently,we analyzed both the metabolomics and transcriptomics data and found that seven metabolites,especially 2’-deoxy-D-ribose(2 d DR),were significantly correlated with differentially expressed genes.In conclusion,our study presents a comprehensive landscape of plasma metabolites in AAA and AS patients,and provides a research direction for pathogenetic mechanisms in atherosclerotic AAA.展开更多
The objective of this experiment was to evaluate the effect of supplementing rumen-protected Lys based on a Lys-deficient diet on liver metabolism in growing Holstein heifers.The experiment was conducted for 3 months ...The objective of this experiment was to evaluate the effect of supplementing rumen-protected Lys based on a Lys-deficient diet on liver metabolism in growing Holstein heifers.The experiment was conducted for 3 months with 36 Holstein heifers(initial body weight:200±9.0 kg;7-month-old).Heifers were randomly assigned to 2 diets based on corn,soybean meal,alfalfa hay,and wheat bran:control,Lysdeficient diet(LD;0.66%Lys in diet),and Lys-adequate diet(LA;1.00%Lys in diet).The results showed no difference in growth performance between the 2 groups(P>0.05).However,there was a clear trend of increasing feed conversion rate with Lys supplementation(0.05<P<0.01).The serum urea nitrogen concentration was significantly decreased,and the aspartate aminotransferase-to-alanine aminotransferase ratio was significantly decreased by Lys supplementation(P<0.05).Moreover,growing heifers fed a Lys-adequate diet had lower levels of urine nitrogen excretion and higher levels of the biological value of nitrogen(P<0.05).Metabolomic analysis revealed that 5 types of phosphatidylcholine and 3 types of ceramide were significantly increased and enriched in sphingolipid metabolism and glycero phospholipid metabolism(P<0.05).His,Leu,and Asp levels were significantly decreased in the liver following Lys supplementation(P<0.05).In conclusion,Lys supplementation may promote the synthesis of body tissue proteins,as evidenced by significantly decreased amino acids in the liver and urine N excretion,it also improves hepatic lipid metabolism by providing lipoprotein precursors.展开更多
Untargeted metabolomics aims to comprehensively profile metabolites as many as possible in biological samples.Recently,ion mobility-mass spectrometry(IM-MS)has emerged as a powerful technology for untargeted metabolom...Untargeted metabolomics aims to comprehensively profile metabolites as many as possible in biological samples.Recently,ion mobility-mass spectrometry(IM-MS)has emerged as a powerful technology for untargeted metabolomics.The emerging role of IM-MS in untargeted metabolomics enables the separation of metabolite isomers and generation of multidimension data to support the identification of metabolites.In this review,we first introduced the basic principles of IM-MS instruments commonly used for untargeted metabolomics.Then,we demonstrated the application of IM-MS for metabolite separation and identification of both known and unknown metabolites.Finally,we discussed the future developments of IM-MS technology to improve untargeted metabolomics.展开更多
桉树因其优异的特性在我国拥有广大的种植面积,在其栽培和生产加工过程会产生大量的桉树皮和桉木屑等副产物,其剩余物资源较丰富。目前已有利用桉木屑栽培食用菌的报道,为了解桉树皮和桉木屑与杂木屑栽培的肺形侧耳(Pleurotus pulmonari...桉树因其优异的特性在我国拥有广大的种植面积,在其栽培和生产加工过程会产生大量的桉树皮和桉木屑等副产物,其剩余物资源较丰富。目前已有利用桉木屑栽培食用菌的报道,为了解桉树皮和桉木屑与杂木屑栽培的肺形侧耳(Pleurotus pulmonarius)代谢物的差异,该研究采用超高效液相色谱-串联质谱(Ultra-High Performance Liquid Chromatography tandem Mass Spectrometry,UHPLC-MS/MS)技术,结合正交偏最小二乘法-判别分析法(Orthogonal Projections to Latent Structures Discriminant Analysis,OPLS-DA)对不同栽培基质条件下肺形侧耳子实体的代谢物进行了非靶向代谢组学研究。以变量权重值(Value Importance in Projection,VIP)>1结合T检验P<0.05为标准进行筛选。结果表明:(1)处理组B(桉木屑)与对照组A(杂木屑)之间有45种差异代谢产物和8种差异代谢物通路,处理组C(桉树皮)与对照组A之间有53种差异代谢产物和15种差异代谢物通路,处理组B与处理组C之间有39种差异代谢产物和5种差异代谢物通路。(2)根据KEGG(Kyoto Encyclopedia of Genes and Genomes)代谢通路富集分析处理组C和对照组A得到4条显著的代谢通路,分别为精氨酸生物合成、精氨酸和脯氨酸代谢、泛酸和辅酶A生物合成和丙氨酸、天冬氨酸和谷氨酸代谢,处理组B和处理组C得到1条显著的代谢通路为组氨酸代谢,而处理组B和对照组A中无显著性代谢通路(P>0.05)。综上表明,利用桉树皮和桉木屑作为肺形侧耳的主要栽培基质(尤其是桉木屑),可以降本增效和实现资源再利用。该研究结果为桉树加工副产物应用于食用菌栽培提供了参考。展开更多
Tartary buckwheat(Fagopyrum tataricum)is an important pseudocereal feed crop with medicinal and nutritional value.Drought is one of the main causes of reduced growth and yield in these plants.We investigated the growt...Tartary buckwheat(Fagopyrum tataricum)is an important pseudocereal feed crop with medicinal and nutritional value.Drought is one of the main causes of reduced growth and yield in these plants.We investigated the growth,physiological,and metabolic responses of the widely promoted Tartary buckwheat variety Chuan Qiao No.1 to polyethylene glycol(PEG)-mediated drought stress.Drought significantly decreased shoot length,shoot biomass and relative water content.Root length,malondialdehyde content,electrolyte leakage,activities of superoxide dismutase,peroxidase,catalase and amylase,and contents of soluble sugar,soluble protein and proline were increased by PEG-mediated drought.Untargeted metabolomics analysis identified 32 core metabolites in seedlings subjected to PEG-mediated drought,16 of which increased—including quercetin,isovitexin,cyanidin 3-O-beta-D-glucoside,L-arginine,and glycerophosphocholine,while the other 16 decreased—including 3-methoxytyramine,2,6-diaminopimelic acid,citric acid,UDP-alpha-D-glucose,adenosine,keto-D-fructose.The 32 core metabolites were enriched in 29 metabolic pathways,including lysine biosynthesis,citrate(TCA)cycle,anthocyanin biosynthesis,and aminoacyl-tRNA biosynthesis.Among them,taurine and hypotaurine metabolism,flavor and flavor biosynthesis,indole alkaline biosynthesis,and alanine,aspartate and glutamate metabolism were the four main metabolic pathways affected by drought.Our findings provide new insights into the physiological and metabolic response mechanisms of Tartary buckwheat to drought stress.展开更多
BACKGROUND Acute lung injury(ALI)and its final severe stage,acute respiratory distress syndrome,are associated with high morbidity and mortality rates in patients due to the lack of effective specific treatments.Gut m...BACKGROUND Acute lung injury(ALI)and its final severe stage,acute respiratory distress syndrome,are associated with high morbidity and mortality rates in patients due to the lack of effective specific treatments.Gut microbiota homeostasis,including that in ALI,is important for human health.Evidence suggests that the gut microbiota improves lung injury through the lung-gut axis.Human umbilical cord mesenchymal cells(HUC-MSCs)have attractive prospects for ALI treatment.This study hypothesized that HUC-MSCs improve ALI via the lung-gut microflora.AIM To explore the effects of HUC-MSCs on lipopolysaccharide(LPS)-induced ALI in mice and the involvement of the lung-gut axis in this process.METHODS C57BL/6 mice were randomly divided into four groups(18 rats per group):Sham,sham+HUC-MSCs,LPS,and LPS+HUC-MSCs.ALI was induced in mice by intraperitoneal injections of LPS(10 mg/kg).After 6 h,mice were intervened with 0.5 mL phosphate buffered saline(PBS)containing 1×10^(6) HUC-MSCs by intraperitoneal injections.For the negative control,100 mL 0.9%NaCl and 0.5 mL PBS were used.Bronchoalveolar lavage fluid(BALF)was obtained from anesthetized mice,and their blood,lungs,ileum,and feces were obtained by an aseptic technique following CO_(2) euthanasia.Wright’s staining,enzyme-linked immunosorbent assay,hematoxylin-eosin staining,Evans blue dye leakage assay,immunohistochemistry,fluorescence in situ hybridization,western blot,16S rDNA sequencing,and non-targeted metabolomics were used to observe the effect of HUC-MSCs on ALI mice,and the involvement of the lung-gut axis in this process was explored.One-way analysis of variance with post-hoc Tukey’s test,independent-sample Student’s t-test,Wilcoxon rank-sum test,and Pearson correlation analysis were used for statistical analyses.RESULTS HUC-MSCs were observed to improve pulmonary edema and lung and ileal injury,and decrease mononuclear cell and neutrophil counts,protein concentrations in BALF and inflammatory cytokine levels in the serum,lung,and ileum of ALI mice.Especially,HUC-MSCs decreased Evans blue concentration and Toll-like receptor 4,myeloid differentiation factor 88,p-nuclear factor kappa-B(NF-κB)/NF-κB,and p-inhibitorαof NF-κB(p-IκBα)/IκBαexpression levels in the lung,and raised the pulmonary vascular endothelial-cadherin,zonula occludens-1(ZO-1),and occludin levels and ileal ZO-1,claudin-1,and occludin expression levels.HUC-MSCs improved gut and BALF microbial homeostases.The number of pathogenic bacteria decreased in the BALF of ALI mice treated with HUCMSCs.Concurrently,the abundances of Oscillospira and Coprococcus in the feces of HUS-MSC-treated ALI mice were significantly increased.In addition,Lactobacillus,Bacteroides,and unidentified_Rikenellaceae genera appeared in both feces and BALF.Moreover,this study performed metabolomic analysis on the lung tissue and identified five upregulated metabolites and 11 downregulated metabolites in the LPS+MSC group compared to the LPS group,which were related to the purine metabolism and the taste transduction signaling pathways.Therefore,an intrinsic link between lung metabolite levels and BALF flora homeostasis was established.CONCLUSION This study suggests that HUM-MSCs attenuate ALI by redefining the gut and lung microbiota.展开更多
Objective:To identify the secondary metabolites of Hei-Yun-Xiang(黑云香),predict its target by network pharmacology,and the target binding activity was demonstrated by molecular docking to explore its mechanism in tre...Objective:To identify the secondary metabolites of Hei-Yun-Xiang(黑云香),predict its target by network pharmacology,and the target binding activity was demonstrated by molecular docking to explore its mechanism in treating inflammatory diseases.Methods:The Untargeted Metabolomics was used to identify the total components of secondary metabolites in Hei-Yun-Xiang.The Hei-Yun-Xiang's active components were screened by the oral bioavailability>30%and drug-likeness>0.10 from the total component of Hei-Yun-Xiang.The TCMSP and standard chemical databases screened the main active components.The Swiss target prediction and the UniProt database predicted the potential targets of Hei-Yun-Xiang's active components.To obtain the potential targets of Hei-Yun-Xiang in the treatment of inflammatory diseases,we compare the above targets with OMIM,NCBI,and GENECARD.Combined with string analysis of Hei-Yun-Xiang's target protein interaction,the R4.0.3 software was used to enrich and analyze Hei-Yun-Xiang's potential targets'GO annotation and KEGG pathway.The"Mongolian medicine-active component-core target-pathway"network structure diagram of Hei-Yun-Xiang in anti-inflammatory effect was constructed using Cytoscape 3.8.0 software.AutoDock Vina 1.1.2 molecular docking software was used to target proteins and the active components interaction model.Results:22 highly active compounds were screened from Hei-Yun-Xiang,of which 19 were related to inflammation.324 targets related to inflammation were predicted and analyzed;targets were mainly composed of genes such as inflammatory biological processes induced by bacterial molecules such as lipopolysaccharide,cell membrane function,and serine/threonine kinase activation.Targets were enriched into the 179 KEGG-related pathways.Pathways mainly include lipid metabolism,arteriosclerosis,neuroactive receptor-ligand binding pathway,PI3K Akt signaling pathway.Molecular docking demonstrated that astrapterocarpan,chimonanthine,columbianetin_acetate,tretinoin,tussilagone,columbianetin_acetate,n-Feruloyltyramine had good binding to eight key proteins,and AKT1 and HSP90AA1 was the target protein with the best binding activity,suggesting that AKT1 and HSP90AA1 could be the essential mediator responsible for signaling transduction after Hei-Yun-Xiang administration.Conclusion:Hei-Yun-Xiang can inhibit the inflammatory reaction by its multi-component,multi-target,and multi-channel treatment of inflammatory disease,interfering with the inflammatory reaction induced by bacterial molecules such as lipopolysaccharide or by intervening in lipid and inflammation-related arteriosclerosis-related pathways.展开更多
文摘Objective:As an important part of metabolomics analysis,untargeted metabolomics has become a powerful tool in the study of tumor mechanisms and the discovery of metabolic markers with high-throughput spectrometric data which also poses great challenges to data analysis,from the extraction of raw data to the identification of differential metabolites.To date,a large number of analytical tools and processes have been developed and constructed to serve untargeted metabolomics research.The different selection of analytical tools and parameter settings lead to varied results of untargeted metabolomics data.Our goal is to establish an easily operated platform and obtain a repeatable analysis result.Methods:We used the R language basic environment to construct the preprocessing system of the original data and the LAMP(Linux+Apache+MySQL+PHP)architecture to build a cloud mass spectrum data analysis system.Results:An open-source analysis software for untargeted metabolomics data(openNAU)was constructed.It includes the extraction of raw mass data and quality control for the identification of differential metabolic ion peaks.A reference metabolomics database based on public databases was also constructed.Conclusions:A complete analysis system platform for untargeted metabolomics was established.This platform provides a complete template interface for the addition and updating of the analysis process,so we can finish complex analyses of untargeted metabolomics with simple human-computer interactions.The source code can be downloaded from https://github.com/zjuRong/openNAU.
基金Supported by the University Scientific Research Projects of Anhui,No. KJ2020A0401 and 2022AH050491the open fund of the Ministry of Education Key Laboratory of Glucolipid Metabolic Disorder,No. GYDKFXM01+5 种基金the Anhui University Collaborative Innovation Project,No. GXXT-2020-025the National Natural Science Foundation of China,No. 82174153the National Key Research and Development Program,No. 2018YFC1704202the Anhui Provincial Quality Engineering Project of Universities,No. 2021jyxm0834the Major and Difficult Diseases Project of Anhui Province,No. 2021zdynjb06the Clinical Research Project of Anhui University of Traditional Chinese Medicine,No. 2021yfylc01
文摘BACKGROUND In recent years,the incidence of type 2 diabetes(T2DM)has shown a rapid growth trend.Goto Kakizaki(GK)rats are a valuable model for the study of T2DM and share common glucose metabolism features with human T2DM patients.A series of studies have indicated that T2DM is associated with the gut microbiota composition and gut metabolites.We aimed to systematically characterize the faecal gut microbes and metabolites of GK rats and analyse the relationship between glucose and insulin resistance.AIM To evaluate the gut microbial and metabolite alterations in GK rat faeces based on metagenomics and untargeted metabolomics.METHODS Ten GK rats(model group)and Wistar rats(control group)were observed for 10 wk,and various glucose-related indexes,mainly including weight,fasting blood glucose(FBG)and insulin levels,homeostasis model assessment of insulin resistance(HOMA-IR)and homeostasis model assessment ofβcell(HOMA-β)were assessed.The faecal gut microbiota was sequenced by metagenomics,and faecal metabolites were analysed by untargeted metabolomics.Multiple metabolic pathways were evaluated based on the differential metabolites identified,and the correlations between blood glucose and the gut microbiota and metabolites were analysed.RESULTS The model group displayed significant differences in weight,FBG and insulin levels,HOMA-IR and HOMA-βindexes(P<0.05,P<0.01)and a shift in the gut microbiota structure compared with the control group.The results demonstrated significantly decreased abundances of Prevotella sp.CAG:604 and Lactobacillus murinus(P<0.05)and a significantly increased abundance of Allobaculum stercoricanis(P<0.01)in the model group.A correlation analysis indicated that FBG and HOMA-IR were positively correlated with Allobaculum stercoricanis and negatively correlated with Lactobacillus murinus.An orthogonal partial least squares discriminant analysis suggested that the faecal metabolic profiles differed between the model and control groups.Fourteen potential metabolic biomarkers,including glycochenodeoxycholic acid,uric acid,13(S)-hydroxyoctadecadienoic acid(HODE),N-acetylaspartate,β-sitostenone,sphinganine,4-pyridoxic acid,and linoleic acid,were identified.Moreover,FBG and HOMA-IR were found to be positively correlated with glutathione,13(S)-HODE,uric acid,4-pyridoxic acid and allantoic acid and negatively correlated with 3-α,7-α,chenodeoxycholic acid glycine conjugate and 26-trihydroxy-5-β-cholestane(P<0.05,P<0.01).Allobaculum stercoricanis was positively correlated with linoleic acid and sphinganine(P<0.01),and 2-methyl-3-hydroxy-5-formylpyridine-4-carboxylate was negatively associated with Prevotella sp.CAG:604(P<0.01).The metabolic pathways showing the largest differences were arginine biosynthesis;primary bile acid biosynthesis;purine metabolism;linoleic acid metabolism;alanine,aspartate and glutamate metabolism;and nitrogen metabolism.CONCLUSION Metagenomics and untargeted metabolomics indicated that disordered compositions of gut microbes and metabolites may be common defects in GK rats.
基金Science and Technology Project Task Book of Beijing,No.Z171100001717008.
文摘BACKGROUND Nonalcoholic fatty liver disease(NAFLD)has become one of the most common chronic liver diseases in the world.In our early clinical data and questionnaire analysis of NAFLD,it was found that the body mass index of some patients did not meet the diagnostic criteria for overweight or obesity.The consumption of high-temperature-processed foods such as fried food,hot pot and barbecue is closely related to the occurrence of nonobese NAFLD.Reducing the intake of this kind of food can reduce disease severity and improve prognosis.AIM To explore the untargeted metabolomics characteristics of nonobese nonalcoholic fatty liver disease in Sprague-Dawley rats induced by high-temperatureprocessed feed.METHODS Fifty-four male Sprague-Dawley rats were divided into three groups:The control group received a standard diet;the nonfried soybeans(NDFS)group received 60%NDFS and 40%basic feed and the dry-fried soybeans(DFS)group received 60%DFS and 40%basic feed.Six rats were sacrificed at week 4,8,and 12 in each group.The food intake,body weight,Lee’s index,liver index,serological index and hepatic histopathology were assessed.Untargeted metabolomics characteristics were used to analyze the changes in liver metabolites of rats at week 12.Correlations between metabolites and pathology scores between the DFS and control groups and between the DFS and NDFS groups were analyzed.We selected some of the metabolites,both within the pathway and outside of the pathway,to explain preliminarily the difference in liver pathology in the three groups of rats.RESULTS There were no statistically significant differences in the food intake,body weight,Lee's index or serological index between the DFS group and the control group(P>0.05).At week 8 and week 12,the steatosis scores in the DFS group were significantly higher than those in the other two groups(P<0.05).At week 12,the liver index of the DFS group was the lowest(NDFS group vs DFS group,P<0.05).The fibrosis score in the DFS group was significantly higher than those in the other two groups(P<0.05).The correlation analysis of the liver pathology score and differential metabolites in the DFS and NDFS groups showed that there were 10 strongly correlated substances:Five positively correlated substances and five negatively correlated substances.The positively correlated substances included taurochenodeoxycholate-3-sulfate,acetylcarnitine,20a,22bdihydroxycholesterol,13E-tetranor-16-carboxy-LTE4 and taurocholic acid.The negatively correlated substances included choline,cholesterane-3,7,12,25-tetrol-3-glucuronide,nicotinamide adenine dinucleotide phosphate,lysoPC[16:1(9Z)]and glycerol 3-phosphate.The correlation analysis of the liver pathology score and differential metabolites in the DFS and control groups showed that there were 13 strongly correlated substances:Four positively correlated substances and 9 negatively correlated substances.The positively correlated substances included 4-hydroxy-6-eicosanone,3-phosphoglyceric acid,13-hydroxy-9-methoxy-10-oxo-11-octadecenoic acid and taurochenodeoxycholate-3-sulfate.The negatively correlated substances included lysoPC[16:1(9Z)],S-(9-hydroxy-PGA1)-glutathione,lysoPC[20:5(5Z,8Z,11Z,14Z,17Z)],SM(d18:1/14:0),nicotinamide adenine dinucleotide phosphate,5,10-methylene-THF,folinic acid,N-lactoylglycine and 6-hydroxy-5-methoxyindole glucuronide.CONCLUSION We successfully induced liver damage in rats by using a specially prepared hightemperature-processed feed and explored the untargeted metabolomics characteristics.
基金National Natural Science Foundation of China(Grant No.81872996)Natural Science Foundation of Tianjin of China(Grant No.20JCYBJC00060).
文摘Quality control of ginseng currently is mainly based on ginsenoside analysis,but rarely focuses on the volatile organic components.In the current work,an untargeted metabolomics approach,by headspace solid-phase micro-extraction gas chromatography/mass spectrometry(HS-SPME-GC/MS),was elaborated and further employed to holistically compare the compositional difference of the volatile components simultaneously from 12 Panax herbal medicines,which included P.ginseng(PG),P.quinquefolius(PQ),P.notoginseng(PN),red ginseng(PGR),P.ginseng leaf(PGL),P.quinquefolius leaf(PQL),P.notoginseng leaf(PNL),P.ginseng flower(PGF),P.quinquefolius flower(PQF),P.notoginseng flower(PNF),P.japonicus(PJ),and P.japonicus var.major(PJvm).Chromatographic separation was performed on an HP-5MS elastic quartz capillary column using helium as the carrier gas,enabling good resolution within 1 h.We were able to characterize totally 259 volatile compounds,including 82 terpenes(T),46 alcohols(Alc),29 ketones(K),25 aldehydes(Ald),21 esters(E),and the others.By analyzing 90 batches of ginseng samples based on the untargeted metabolomics workflows,236 differential ions were unveiled,and accordingly 36 differential volatile components were discovered.It is the first report that simultaneously compares the compositional difference of volatile components among 12 Panax herbal medicines,and useful information is provided for the quality control of ginseng aside from the well-known ginsenosides.
文摘Polymerase-tautomeric model for untargeted delayed base substitution mutations is proposed.Structural analysis of bases insertion showed that any canonical bases may be inserted opposite rare tautomeric forms of thymine T3*,adenines A2*and A4*so that between them hydrogen bonds are formed.Canonical adenine and cytosine can be incorporated opposite canonical thymine only.Canonical thymine and guanine can be incorporated opposite canonical adenine only.If in the synthesis of DNA containing rare tautomeric forms of thymine T3*,adenines A2*and A4*,involved DNA polymerases with relatively high fidelity of synthesis,mutations not appear.However,if further DNA synthesis will involve DNA polymerases having a low fidelity of synthesis,there may be base substitution mutations.It was shown that the conclusion made in the Tomasetti and Vogelstein cancer risk model that the formation of about 67%of all mutations was not caused by exposure to any mutagens is erroneous.
基金Supported by the Scientific Foundation of Administration of Traditional Chinese Medicine of Hebei Province,China,No.2023257.
文摘BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therapeutic effects and the possible mechanism of JPGS on DN.METHODS We first evaluated the therapeutic potential of JPGS on a DN mouse model.We then investigated the effect of JPGS on the renal metabolite levels of DN mice using non-targeted metabolomics.Furthermore,we examined the effects of JPGS on c-Jun N-terminal kinase(JNK)/P38-mediated apoptosis and the inflammatory responses mediated by toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3).RESULTS The ameliorative effects of JPGS on DN mice included the alleviation of renal injury and the control of inflammation and oxidative stress.Untargeted metabolomic analysis revealed that JPGS altered the metabolites of the kidneys in DN mice.A total of 51 differential metabolites were screened.Pathway analysis results indicated that nine pathways significantly changed between the control and model groups,while six pathways significantly altered between the model and JPGS groups.Pathways related to cysteine and methionine metabolism;alanine,tryptophan metabolism;aspartate and glutamate metabolism;and riboflavin metabolism were identified as the key pathways through which JPGS affects DN.Further experimental validation showed that JPGS treatment reduced the expression of TLR4/NF-κB/NLRP3 pathways and JNK/P38 pathway-mediated apoptosis related factors.CONCLUSION JPGS could markedly treat mice with streptozotocin(STZ)-induced DN,which is possibly related to the regulation of several metabolic pathways found in kidneys.Furthermore,JPGS could improve kidney inflammatory responses and ameliorate kidney injuries in DN mice via the TLR4/NF-κB/NLRP3 pathway and inhibit JNK/P38 pathwaymediated apoptosis in DN mice.
文摘In the present study, total membrane proteins from tumor cell lines including HepG2, Hep3 B2,H226, Ovcar3 and N87 were extracted and digested with γLys C and trypsin. The resulting peptide lysate were pre-fractionated and subjected to untargeted quantitative proteomics analysis using a high resolution mass spectrometer. The mass spectra were processed by the Max Quant and the protein abundances were estimated using total peak area(TPA) method. A total of 6037 proteins were identified, and the analysis resulted in the identification of 2647 membrane proteins. Of those, tumor antigens and absorption,metabolism, disposition and elimination(ADME) proteins including UDP-glucuronosyltransferase,cytochrome P450, solute carriers and ATP-binding cassette transporters were detected and disclosed significant variations among the cell lines. The principal component analysis was performed for the cluster of cell lines. The results demonstrated that H226 is closely related with N87, while Hep3 B2 aligned with HepG2. The protein cluster of Ovcar3 was apart from that of other cell lines investigated. By providing for the first time quantitative untargeted proteomics analysis, the results delineated the expression profiles of membrane proteins. These findings provided a useful resource for selecting targets of choice for anticancer therapy through advancing data obtained from preclinical tumor cell line models to clinical outcomes.
基金supported by the National Natural Science Foundation of China(Nos.51890894,81770481,and 82070492)the Chinese Academy of Medical SciencesInnovation Fund for Medical Sciences(CIFMS 2017-I2M-1-008)。
文摘Abdominal aortic aneurysm(AAA)and atherosclerosis(AS)have considerable similarities in clinical risk factors and molecular pathogenesis.The aim of our study was to investigate the differences between AAA and AS from the perspective of metabolomics,and to explore the potential mechanisms of differential metabolites via integration analysis with transcriptomics.Plasma samples from 32 AAA and 32 AS patients were applied to characterize the metabolite profiles using untargeted liquid chromatography-mass spectrometry(LC-MS).A total of 18 remarkably different metabolites were identified,and a combination of seven metabolites could potentially serve as a biomarker to distinguish AAA and AS,with an area under the curve(AUC)of0.93.Subsequently,we analyzed both the metabolomics and transcriptomics data and found that seven metabolites,especially 2’-deoxy-D-ribose(2 d DR),were significantly correlated with differentially expressed genes.In conclusion,our study presents a comprehensive landscape of plasma metabolites in AAA and AS patients,and provides a research direction for pathogenetic mechanisms in atherosclerotic AAA.
基金supported,in part,by the Key Research and Development Program of Hebei Province(19226621D)Earmarked Fund for Beijing Dairy Industry Innovation Consortium of Agriculture Research System,Chinese Academy of Agricultural Science and Technology Innovation Project(CAAS-XTCX-2016011-01)+2 种基金Fundamental Research Funds for Central Non-profit Scientific Institution(Y2019CG08)Agricultural Science and Technology Innovation Program(CAAS-ASTIP-2017-FRI-04)Science and Technology Open Cooperation Project of Henan Province(182106000035)Study on the Pattern of Diet Amino Acid for Different Physiological Stages of Heifers。
文摘The objective of this experiment was to evaluate the effect of supplementing rumen-protected Lys based on a Lys-deficient diet on liver metabolism in growing Holstein heifers.The experiment was conducted for 3 months with 36 Holstein heifers(initial body weight:200±9.0 kg;7-month-old).Heifers were randomly assigned to 2 diets based on corn,soybean meal,alfalfa hay,and wheat bran:control,Lysdeficient diet(LD;0.66%Lys in diet),and Lys-adequate diet(LA;1.00%Lys in diet).The results showed no difference in growth performance between the 2 groups(P>0.05).However,there was a clear trend of increasing feed conversion rate with Lys supplementation(0.05<P<0.01).The serum urea nitrogen concentration was significantly decreased,and the aspartate aminotransferase-to-alanine aminotransferase ratio was significantly decreased by Lys supplementation(P<0.05).Moreover,growing heifers fed a Lys-adequate diet had lower levels of urine nitrogen excretion and higher levels of the biological value of nitrogen(P<0.05).Metabolomic analysis revealed that 5 types of phosphatidylcholine and 3 types of ceramide were significantly increased and enriched in sphingolipid metabolism and glycero phospholipid metabolism(P<0.05).His,Leu,and Asp levels were significantly decreased in the liver following Lys supplementation(P<0.05).In conclusion,Lys supplementation may promote the synthesis of body tissue proteins,as evidenced by significantly decreased amino acids in the liver and urine N excretion,it also improves hepatic lipid metabolism by providing lipoprotein precursors.
基金The work was supported by National Natural Science Foundation of China(Grant No.31971356)Shang-hai Municipal Science and Technology Major Project(Grant No.2019SHZDZX02)。
文摘Untargeted metabolomics aims to comprehensively profile metabolites as many as possible in biological samples.Recently,ion mobility-mass spectrometry(IM-MS)has emerged as a powerful technology for untargeted metabolomics.The emerging role of IM-MS in untargeted metabolomics enables the separation of metabolite isomers and generation of multidimension data to support the identification of metabolites.In this review,we first introduced the basic principles of IM-MS instruments commonly used for untargeted metabolomics.Then,we demonstrated the application of IM-MS for metabolite separation and identification of both known and unknown metabolites.Finally,we discussed the future developments of IM-MS technology to improve untargeted metabolomics.
文摘桉树因其优异的特性在我国拥有广大的种植面积,在其栽培和生产加工过程会产生大量的桉树皮和桉木屑等副产物,其剩余物资源较丰富。目前已有利用桉木屑栽培食用菌的报道,为了解桉树皮和桉木屑与杂木屑栽培的肺形侧耳(Pleurotus pulmonarius)代谢物的差异,该研究采用超高效液相色谱-串联质谱(Ultra-High Performance Liquid Chromatography tandem Mass Spectrometry,UHPLC-MS/MS)技术,结合正交偏最小二乘法-判别分析法(Orthogonal Projections to Latent Structures Discriminant Analysis,OPLS-DA)对不同栽培基质条件下肺形侧耳子实体的代谢物进行了非靶向代谢组学研究。以变量权重值(Value Importance in Projection,VIP)>1结合T检验P<0.05为标准进行筛选。结果表明:(1)处理组B(桉木屑)与对照组A(杂木屑)之间有45种差异代谢产物和8种差异代谢物通路,处理组C(桉树皮)与对照组A之间有53种差异代谢产物和15种差异代谢物通路,处理组B与处理组C之间有39种差异代谢产物和5种差异代谢物通路。(2)根据KEGG(Kyoto Encyclopedia of Genes and Genomes)代谢通路富集分析处理组C和对照组A得到4条显著的代谢通路,分别为精氨酸生物合成、精氨酸和脯氨酸代谢、泛酸和辅酶A生物合成和丙氨酸、天冬氨酸和谷氨酸代谢,处理组B和处理组C得到1条显著的代谢通路为组氨酸代谢,而处理组B和对照组A中无显著性代谢通路(P>0.05)。综上表明,利用桉树皮和桉木屑作为肺形侧耳的主要栽培基质(尤其是桉木屑),可以降本增效和实现资源再利用。该研究结果为桉树加工副产物应用于食用菌栽培提供了参考。
基金We acknowledge the Project of National Key Research and Development Program of China(2020YFD1001403)China Agriculture Research System(CARS-07-B-1)+3 种基金Science&Technology Department of Sichuan Province(2022YFQ0041)the National Natural Science Foundation of China(31601260,32160428)Innovative Training Program for College Students(S202111079058)Special Research Fund from Key Laboratory of Coarse Cereal Processing,Ministry of Agriculture and Rural Affairs(2020CC012)to facilitate the research.
文摘Tartary buckwheat(Fagopyrum tataricum)is an important pseudocereal feed crop with medicinal and nutritional value.Drought is one of the main causes of reduced growth and yield in these plants.We investigated the growth,physiological,and metabolic responses of the widely promoted Tartary buckwheat variety Chuan Qiao No.1 to polyethylene glycol(PEG)-mediated drought stress.Drought significantly decreased shoot length,shoot biomass and relative water content.Root length,malondialdehyde content,electrolyte leakage,activities of superoxide dismutase,peroxidase,catalase and amylase,and contents of soluble sugar,soluble protein and proline were increased by PEG-mediated drought.Untargeted metabolomics analysis identified 32 core metabolites in seedlings subjected to PEG-mediated drought,16 of which increased—including quercetin,isovitexin,cyanidin 3-O-beta-D-glucoside,L-arginine,and glycerophosphocholine,while the other 16 decreased—including 3-methoxytyramine,2,6-diaminopimelic acid,citric acid,UDP-alpha-D-glucose,adenosine,keto-D-fructose.The 32 core metabolites were enriched in 29 metabolic pathways,including lysine biosynthesis,citrate(TCA)cycle,anthocyanin biosynthesis,and aminoacyl-tRNA biosynthesis.Among them,taurine and hypotaurine metabolism,flavor and flavor biosynthesis,indole alkaline biosynthesis,and alanine,aspartate and glutamate metabolism were the four main metabolic pathways affected by drought.Our findings provide new insights into the physiological and metabolic response mechanisms of Tartary buckwheat to drought stress.
基金the Key Research and Development Project of Science and Technology Department of Zhejiang Province,No.2019C03041.
文摘BACKGROUND Acute lung injury(ALI)and its final severe stage,acute respiratory distress syndrome,are associated with high morbidity and mortality rates in patients due to the lack of effective specific treatments.Gut microbiota homeostasis,including that in ALI,is important for human health.Evidence suggests that the gut microbiota improves lung injury through the lung-gut axis.Human umbilical cord mesenchymal cells(HUC-MSCs)have attractive prospects for ALI treatment.This study hypothesized that HUC-MSCs improve ALI via the lung-gut microflora.AIM To explore the effects of HUC-MSCs on lipopolysaccharide(LPS)-induced ALI in mice and the involvement of the lung-gut axis in this process.METHODS C57BL/6 mice were randomly divided into four groups(18 rats per group):Sham,sham+HUC-MSCs,LPS,and LPS+HUC-MSCs.ALI was induced in mice by intraperitoneal injections of LPS(10 mg/kg).After 6 h,mice were intervened with 0.5 mL phosphate buffered saline(PBS)containing 1×10^(6) HUC-MSCs by intraperitoneal injections.For the negative control,100 mL 0.9%NaCl and 0.5 mL PBS were used.Bronchoalveolar lavage fluid(BALF)was obtained from anesthetized mice,and their blood,lungs,ileum,and feces were obtained by an aseptic technique following CO_(2) euthanasia.Wright’s staining,enzyme-linked immunosorbent assay,hematoxylin-eosin staining,Evans blue dye leakage assay,immunohistochemistry,fluorescence in situ hybridization,western blot,16S rDNA sequencing,and non-targeted metabolomics were used to observe the effect of HUC-MSCs on ALI mice,and the involvement of the lung-gut axis in this process was explored.One-way analysis of variance with post-hoc Tukey’s test,independent-sample Student’s t-test,Wilcoxon rank-sum test,and Pearson correlation analysis were used for statistical analyses.RESULTS HUC-MSCs were observed to improve pulmonary edema and lung and ileal injury,and decrease mononuclear cell and neutrophil counts,protein concentrations in BALF and inflammatory cytokine levels in the serum,lung,and ileum of ALI mice.Especially,HUC-MSCs decreased Evans blue concentration and Toll-like receptor 4,myeloid differentiation factor 88,p-nuclear factor kappa-B(NF-κB)/NF-κB,and p-inhibitorαof NF-κB(p-IκBα)/IκBαexpression levels in the lung,and raised the pulmonary vascular endothelial-cadherin,zonula occludens-1(ZO-1),and occludin levels and ileal ZO-1,claudin-1,and occludin expression levels.HUC-MSCs improved gut and BALF microbial homeostases.The number of pathogenic bacteria decreased in the BALF of ALI mice treated with HUCMSCs.Concurrently,the abundances of Oscillospira and Coprococcus in the feces of HUS-MSC-treated ALI mice were significantly increased.In addition,Lactobacillus,Bacteroides,and unidentified_Rikenellaceae genera appeared in both feces and BALF.Moreover,this study performed metabolomic analysis on the lung tissue and identified five upregulated metabolites and 11 downregulated metabolites in the LPS+MSC group compared to the LPS group,which were related to the purine metabolism and the taste transduction signaling pathways.Therefore,an intrinsic link between lung metabolite levels and BALF flora homeostasis was established.CONCLUSION This study suggests that HUM-MSCs attenuate ALI by redefining the gut and lung microbiota.
基金The Inner Mongolia Autonomous Region Personalized Medicine Engineering Technology Research Center Open Foundation(MDK2021056,MDK2019083,MDK2019085)The Central Government Guiding Special Funds for Development of Local Science and Technology(2020ZY0020)。
文摘Objective:To identify the secondary metabolites of Hei-Yun-Xiang(黑云香),predict its target by network pharmacology,and the target binding activity was demonstrated by molecular docking to explore its mechanism in treating inflammatory diseases.Methods:The Untargeted Metabolomics was used to identify the total components of secondary metabolites in Hei-Yun-Xiang.The Hei-Yun-Xiang's active components were screened by the oral bioavailability>30%and drug-likeness>0.10 from the total component of Hei-Yun-Xiang.The TCMSP and standard chemical databases screened the main active components.The Swiss target prediction and the UniProt database predicted the potential targets of Hei-Yun-Xiang's active components.To obtain the potential targets of Hei-Yun-Xiang in the treatment of inflammatory diseases,we compare the above targets with OMIM,NCBI,and GENECARD.Combined with string analysis of Hei-Yun-Xiang's target protein interaction,the R4.0.3 software was used to enrich and analyze Hei-Yun-Xiang's potential targets'GO annotation and KEGG pathway.The"Mongolian medicine-active component-core target-pathway"network structure diagram of Hei-Yun-Xiang in anti-inflammatory effect was constructed using Cytoscape 3.8.0 software.AutoDock Vina 1.1.2 molecular docking software was used to target proteins and the active components interaction model.Results:22 highly active compounds were screened from Hei-Yun-Xiang,of which 19 were related to inflammation.324 targets related to inflammation were predicted and analyzed;targets were mainly composed of genes such as inflammatory biological processes induced by bacterial molecules such as lipopolysaccharide,cell membrane function,and serine/threonine kinase activation.Targets were enriched into the 179 KEGG-related pathways.Pathways mainly include lipid metabolism,arteriosclerosis,neuroactive receptor-ligand binding pathway,PI3K Akt signaling pathway.Molecular docking demonstrated that astrapterocarpan,chimonanthine,columbianetin_acetate,tretinoin,tussilagone,columbianetin_acetate,n-Feruloyltyramine had good binding to eight key proteins,and AKT1 and HSP90AA1 was the target protein with the best binding activity,suggesting that AKT1 and HSP90AA1 could be the essential mediator responsible for signaling transduction after Hei-Yun-Xiang administration.Conclusion:Hei-Yun-Xiang can inhibit the inflammatory reaction by its multi-component,multi-target,and multi-channel treatment of inflammatory disease,interfering with the inflammatory reaction induced by bacterial molecules such as lipopolysaccharide or by intervening in lipid and inflammation-related arteriosclerosis-related pathways.
