Effects of methimazole on urinary metabolomics in hyperthyroidism rats / 中国药房

OBJECTIVE To study the effects of methimazole on the urinary metabolomics of hyperthyroidism rats， and to preliminarily investigate its possible mechanism. METHODS Thirty SD rats were randomly divided into control group， model group and methimazole group， with 10 rats in each group. Except for the control group， the rats in the other two groups were given Levothyroxine sodium tablets 160 mg/kg by intragastric administration for 15 days； at the same time， methimazole group was additionally given methimazole 3.6 mg/kg daily by intragastric administration every day. The basic condition of the rats was observed， and the body weight and anal temperature were measured. After the last medication， the serum levels of triiodothyronine （T3）， tetraiodothyronine （T4）， free triiodothyronine （FT3）， free tetraiodothyronine （FT4）， and thyroid stimulating hormone （TSH） were determined； 24-hour urine was collected on the 15th day after administration. UPLC-TOF-MS was used to analyze the urine metabolomics of rats. Principal component analysis and orthogonal partial least squares-discriminant analysis were used to screen out related differential metabolites， and potential metabolic pathways were analyzed by using HMDB and KEGG. RESULTS Compared with the control group， the rectal temperature， serum levels of T3， T4， FT3 and FT4， the expressions of differential metabolites sebacic acid， cholic acid 3-O-glucuronic acid and N6， N6， N6-trimethyl-L-lysine in urine were significantly up-regulated， while body weight， serum level of TSH， the expressions of deoxycytidine and 2-oxo-4-methylthiobutanoic acid in urine were significantly down-regulated （P＜0.01）. Compared with model group， above indexes of rats were reversed significantly in methimazole group （P＜0.01 or P＜0.05）. Above five differential metabolites were mainly involved in four signaling pathways: pentose and glucuronate interaction， lysine degradation， cysteine and methionine metabolism， and pyrimidine metabolism. CONCLUSIONS Methimazole might improve hyperthyroidism by modulating the four pathways of pentose and glucuronate interaction， lysine degradation， cysteine and methionine metabolism， and pyrimidine metabolism.

Metabolomics analysis of serum and urine in patients with traumatic spinal cord injury / 中国组织工程研究

BACKGROUND:Traumatic spinal cord injury primarily relies on scale assessment and imaging examinations in clinical practice.However,there are limitations in predicting the prognosis of the injury.Therefore,the use of metabolomics technology for biomarker screening is significant for estimating the extent of damage,injury and recovery,as well as developing new therapies. OBJECTIVE:To characterize the metabolic features of patients with traumatic spinal cord injury using metabolomics technology and explore potential biomarkers and disrupted metabolic pathways. METHODS:Serum and urine samples were collected from 20 patients with traumatic spinal cord injury(observation group)and 10 healthy subjects(control group).Metabolites were analyzed and multivariate statistical analysis was then performed for data processing to screen differential metabolites.Metabolic pathway enrichment was performed using MetaboAnalyst software.Logistic regression was applied to construct a biomarker combination model,and its relationship with the American Spinal Injury Association grading was analyzed. RESULTS AND CONCLUSION:Significant differences in 160 and 73 metabolites were detected in the serum and urine samples of the two groups,respectively.Pathway enrichment analysis showed evident disturbances in lipid metabolism after traumatic spinal cord injury,including sphingolipid,arachidonic acid,α-linolenic acid,and arachidonic acid metabolism,as well as glycerophospholipid and inositol phosphate biosynthesis.The combination of two identified biomarkers,telmisartan and quercetin glycoside,showed a correlation with the American Spinal Injury Association grading in both serum and urine levels.Thus,metabolomics technology provides assistance in further understanding the pathological mechanisms of traumatic spinal cord injury and screening therapeutic targets.The identified metabolic biomarker combination may serve as a reference for assessing the severity of traumatic spinal cord injury.

Serum metabolomics-based study on the mechanism of action of bergapten in the treatment of liver fibrosis / 中国药房

OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

Analysis of Suitable Processing Time of Rehmanniae Radix Praeparata Processed with Amomi Fructus and Citri Reticulatae Pericarpium Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

ObjectiveTo investigate the relative content changes of differential metabolites and reducing sugars during the processing process of Rehmanniae Radix Praeparata （RRP） processed with Amomi Fructus （AF） and Citri Reticulatae Pericarpium （CRP）， and to lay the foundation for revealing the processing principle of this characteristic variety. MethodThe samples of the 0-54 h processing process of RRP processed with AF and CRP were taken as the research object， and their secondary metabolites were detected by ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS）. The 0.1% formic acid aqueous solution （A）-acetonitrile （B） was used as the mobile phase for gradient elution （0-1 min， 1%-3%B； 1-10 min， 3%-9%B； 10-15 min， 9%-12%B； 15-22 min， 12%-18%B； 22-31 min， 18%-24%B； 31-35 min， 24%-100%B； 35-36 min， 100%-5%B； 36-40 min， 5%-1%B； 40-45 min， 1%B）， column temperature was 40 ℃， injection volume was 3 μL， flow rate was 0.3 mL·min-1. Electrospray ionization （ESI） was used to scan and collect MS data in the negative ion mode， the scanning range was m/z 50-1 250. Data analysis was carried out using PeakView 1.2 software， and the chemical composition of RRP processed with AF and CRP was identified by combining the literature information and chemical composition databases. The MS data were normalized by MarkerView 1.2， and then the multivariate statistical analysis was applied to screen the differential metabolites， and the changes of the relative contents of the differential metabolites with different processing times was analyzed， finally， correlation analysis was performed between the differential metabolites， the change of the reducing sugar content was combined to determine the most suitable processing time of RRP processed with AF and CRP. ResultA total of 121 compounds were identified from RRP processed with AF and CRP at different processing times， and 12 differential metabolites were screened out by multivariate statistical analysis， including catalpol， hesperidin， isoacteoside， acteoside， narirutin， echinacoside， isomartynoside， decaffeoylacteoside， 6-O-E-feruloylajugol， dihydroxy-7-O-neohesperidin， jionoside D， and rehmapicroside. With the prolongation of processing time， the relative contents of these 12 differential metabolites and reducing sugars changed slightly at 52-54 h. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents of RRP processed with AF and CRP at different processing times， and the suitable processing time of 52-54 h is determined according to the content changes of different metabolites and reducing sugars， which provides a basis for revealing the scientific connotation of the processing principle of this variety.

Metabolomics insights into the mechanism by which Jinshui Chenfei formula ameliorates silica-induced fibrosis in rats / 中国中西医结合急救杂志

Objective To exploring the mechanism of Jinshui Chenfei formula(JCF)in ameliorating silica(SiO2)-induced silicosis fibrosis based on endogenous metabolite changes.Methods A total of 32 SPF male Sprague-Dawley(SD)rats were divided into normal control group,model group,JCF group(9.72 g·kg-1·d-1),and Tetrandrine group(27 mg·kg-1·d-1)according to random number table method.The experimental silicosis model was established by intratracheal injection with SiO2 suspension(250 mg/kg)on day 1.From week 5-8,silicosis rats were treated with tetrandrine or JCF.On the end of week 8,the changes of pulmonary function index,including forced vital capacity(FVC),tidal volume(TV)and lung dynamic compliance(Cydn)were detected.The pathological changes of lung tissue were analyzed by hematoxyline-osin(HE)staining and Masson staining,the severity of focal alveolitis and fibrosis was also evaluated using the Szapiel scale and the Ashcroft scale,the positive staining of collagen Ⅰ(COL Ⅰ)and COL Ⅲ was detected using immunohistochemistry;the protein expression of transforming growth factor-β1(TGF-β1),fibronectin(FN),andα-smooth muscle actin(α-SMA)were measured by Western blotting.The rat serum samples were further screened for differential metabolites using ultra performance liquid chromatographytandem quadrupole time of flight mass spectrometr(UPLC-Q-TOF-MS)and pathway analysis was performed based on MetaboAnalyst 5.0.Results Compared with those in the normal control group,pathological changes such as alveolar structure destruction,the fibrous nodules encapsulated SiO2 particles were increased in lung tissues of rats in model group,alveolitis score and pulmonary fibrosis score were significantly higher(alveolitis score:2.62±0.27 vs.0.20±0.15,pulmonary fibrosis score:5.42±0.66 vs.0.50±0.84,both P<0.01);pulmonary function index including Cydn,FVC,and TV were significantly decreased[Cdyn(mL/cmH2O):0.26±0.03 vs.0.33±0.03,FVC(mL):8.09±0.47 vs.10.99±0.38,TV(mL):1.95±0.19 vs.2.53±0.26,all P<0.01];positive staining of COL Ⅰ,COL Ⅲ and ɑ-SMA,FN,TGF-β1 proteins expression showed higher in lung tissues[positive staining of COL Ⅰ(A value):13.47±1.76 vs.5.77±0.45;positive staining of COL Ⅲ(A value):10.39±0.47 vs.6.19±0.77,FN protein expression(FN/GAPDH):0.33±0.02 vs.0.21±0.07,α-SMA protein expression(α-SMA/GAPDH):1.78±0.16 vs.1.11±0.24,TGF-β1 protein expression(TGF-β1/GAPDH):0.52±0.10 vs.0.11±0.46,all P<0.01].Compared with the model group,the pathological changes of lung tissues were almost restored,alveolitis score and lung fibrosis score were significantly reduced in JCF and Tetrandrine groups(alveolitis score:1.10±0.15,1.33±0.31 vs.2.62±0.27,pulmonary fibrosis score:3.50±0.45,4.33±0.98 vs.5.42±0.66,all P<0.01);the pulmonary function index Cydn,FVC and TV were significantly increased[Cdyn(mL/cmH2O):0.32±0.05,0.31±0.04 vs.0.26±0.03,FVC(mL):9.41±0.85,8.70±0.92 vs.8.09±0.47,TV(mL):2.70±0.19,2.27±0.15 vs.1.95±0.19,all P<0.05];positive staining of COL Ⅰ,COL Ⅲ,and protein expression of FN,ɑ-SMA,and TGF-β1 in lung tissues was significantly decreased[COL Ⅰ(A value):7.09±0.67,8.13±0.64 vs.13.47±1.76,COL Ⅲ(A value):8.19±0.66,8.52±0.22 vs.10.39±0.47,FN protein expression(FN/GAPDH):0.19±0.06,0.24±0.03 vs.0.33±0.02,α-SMA protein expression(α-SMA/GAPDH):0.89±0.41,0.88±0.08 vs.1.78±0.16,TGF-β1 protein expression(TGF-β1/GAPDH):0.04±0.03,0.06±0.01 vs.0.52±0.10,all P<0.05].Metabolomics analysis showed that a total of 10 major differential metabolites were identified between normal control group,model group and JCF group,including arachidonic acid,palmitic acid,indole-3-acetic acid,propionylcarnitine,(S)-4-hydroxymandelonitrile,nalidixic acid,benzocaine,gramine,4-ethylphenol,N-benzylfor mamide.The differential metabolites in silicosis rats reversed by JCF treatment were mainly enriched,including unsaturated fatty acid biosynthesis,arachidonic acid metabolism,tryptophan metabolism,fatty acid elongation,fatty acid degradation and biosynthesis.Conclusion JCF could effectively improve the silicosis fibrosis,which is mainly related to biosynthesis of unsaturated fatty acids biosynthesis,arachidonic acid metabolism,tryptophan metabolism,fatty acid elongation,fatty acid degradation and biosynthesis.

Effects of lentinan on dendritic cell metabolism / 中国药理学通报

Aim To study the effects of lentinan(LNT)on the metabolism of dendritic cells(DCs)by metabonomics, and uncover the potential mechanism of its regulation of DC function. Methods DC2.4 cells were co-incubated with LNT for 24 h, and the activity of the cells was detected by thiazolyl blue tetrazolium bromide(MTT)assay. The contents of interleukin-6(IL-6), tumor necrosis factorα(TNF-α)and interleukin-12(IL-12)in supernatant were detected by enzyme-linked immunosorbent assay(ELISA). The metabolic general changes of DC2.4 cells were detected by Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS), and the differential metabolites were analyzed by multi-distance covariates and bioinformatics, partial least squares-discriminant analysis(PLS-DA). Finally, metabolic pathway analysis was performed by MetaboAnalyst 5.0. Results LNT did not significantly inhibit the activity of DC2.4 cells at the dose of 25100 mg·L-1. LNT(100 mg·L-1)could significantly stimulate the secretion of IL-6, TNF-α and IL-12 in DC2.4 cells. 20 differential metabolites were identified in DC2.4 cells after being stimulated by LNT(100 mg·L-1), which involved 25 metabolic pathways including urea cycle, arginine and proline metabolism. Conclusion The regulation of LNT on DC function involves a variety of amino acid metabolism.

Curative effect of fuzheng xiaoliu granules in the treatment of primary liver cancer and analysis of serum metabolomics / 中国临床药理学与治疗学

AIM: To observe the clinical efficacy of Fuzheng Xiaoliu Granules in the treatment of stage II primary liver cancer and to explore its mechanism of action from the perspective of metabolomics. METHODS: Sixty patients with stage II primary liver cancer who achieved complete remission (CR) after comprehensive interventional therapy were randomly divided into treatment group and placebo group, with 30 patients in each group. They were treated for one year and observed for one year. The one-year recurrence rate, traditional chinese medicine (TCM) syndrome score, alpha-fetoprotein and child-pugh grade were compared between the two groups. The serum metabolites of the two groups before and after treatment were screened by ultra-high liquid chromatography-mass spectrometry technology, and the metabolic pathways and related biological pathways were analyzed. RESULTS: The one-year recurrence rate of the treatment group was significantly lower than that of the placebo group, and the overall improvement rate of TCM syndrome score was significantly better than that of the placebo group (P0.05). Metabolomics results showed that there were 39 and 33 different metabolites in the treatment group before and after treatment and in the two groups after treatment, respectively. After enrichment analysis and topological analysis of the different metabolites, it was found that Fuzheng Xiaoliu Granules could affect amino acid metabolism, fatty acid metabolism and purine metabolism and other metabolic pathways. Before and after treatment in the treatment group and after treatment in the two groups, there were the same differential metabolites and metabolic pathways in the two comparison results. The same differential metabolites with FOLD CHANGE>1 include Stearic acid, Hypoxanthine, Kynurenic acid, Arachidonic acid, and N-Arachidonoyl Dopamine. The same metabolic pathways with Impact>0.1 include Arachidonic acid metabolism and Histidine metabolism. CONCLUSION: Fuzheng xiaoliu granules can effectively reduce the recurrence rate of stage II liver cancer patients after comprehensive intervention and improve the TCM syndrome. It may inhibit the activation of PI3K/Akt and ERK signaling pathways by regulating the content of metabolites involved in metabolic pathways such as amino acids and fatty acids, thereby delaying tumor recurrence.

Dual metabolic platform to analyze differential metabolites in hepatitis B virus-related liver cirrhosis patients with pre-sarcopenia / 中华临床营养杂志

Objective:This study aimed to analyze differential metabolites in patients using a dual metabolic platform and to orientate early nutritional intervention in patients with cirrhosis.Methods:The skeletal muscle index (SMI) was calculated based on computed tomography (CT) measurements of skeletal muscle cross-sectional area at the third lumbar vertebra level. Pre-sarcopenia was diagnosed for males with SMI < 46.96 and for females with SMI < 32.46. Fifteen HBV-related liver cirrhosis patients with pre-sarcopenia were included as Group S while fourteen liver cirrhosis without pre-sarcopenia were Group NS. Liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) analyses were used to detect differential metabolites and disturbed pathways in the two groups.Results:Five pathways and twenty-eight pathways were defined as disturbed pathways in the plasma of liver cirrhosis patients with pre-sarcopenia by LC-MS and GC-MS, respectively. Most of these pathways are related to amino acid metabolism. Forty-two differential metabolites were imported into the disturbed pathways. Moreover, 3-hydroxypropanal, hydrocinnamic acid, betaine aldehyde, phosphohydroxypyruvic acid, (r)-3-hydroxybutyric acid, and creatinine were identified as potential biomarkers for pre-sarcopenia in HBV-related liver cirrhosis.Conclusions:The study identified a total of 33 pathways and related differential metabolites that were disturbed in HBV-related liver cirrhosis with pre-sarcopenia. The amino acid metabolism, urea cycle, and glyoxylate and dicarboxylate metabolism pathways may be associated with pre-sarcopenia in patients with HBV-related liver cirrhosis. These results provide a direction for nutritional supplementation in liver cirrhosis.

Biomarkers of zinc deficiency in left-behind children under 1 year old in Zunyi area / 中华实用儿科临床杂志

Objective:To analyze the urine of normal healthy left-behind children under 1 year old and left-behind children with zinc deficiency under 1 year old in Zunyi area using hydrogen nuclear magnetic resonance ( 1HNMR), thus providing a new biomarker for the early diagnosis of zinc deficiency. Methods:From January to August 2018, a total of 40 normal healthy left-behind children under 1 year old in Zunyi area(healthy control group)[22 males and 18 females, average age of (7.78±3.62) months, average height of (65.01±2.67) cm and average body mass of (7.15±1.59) kg] and 40 age-matched left-behind children with zinc deficiency in the same region(zinc deficiency group)[19 males and 21 females, average age of (7.89±3.57) months, average height of (64.25±2.95) cm and average body mass of (7.02±1.68) kg] were included for a cross-sectional study by stratified sampling.The urine 1HNMR spectra of children in the 2 groups were measured, and the age, height, body mass and serum zinc content of children in the 2 groups were compared.The metabolites of the 2 groups were compared by metabono-mics technology combined with multivariate statistical analysis, and the differential metabolites of children with zinc deficiency were screened out. Results:There were no significant differences in age, height and body mass between the 2 groups (all P>0.05). The serum zinc level of healthy control group was significantly higher than that of zinc deficiency group [(54.3±3.06) mmol/L vs.(39.2±3.77) mmol/L, t=22.65, P<0.05]. Urine 1HNMR spectrogram results showed that compared with healthy controls, 4-hydroxyphenylpyruvic acid, phenyl acetyl glycine, and hippuric acid salt water were significantly lower in zinc deficiency group ( r=-0.620, -0.689, and -0.721, respectively, all | r|>0.602, all P<0.05). Conclusions:Zinc deficiency in left-behind children under 1 year old in Zunyi area is mainly manifested by decreased metabolites of 4-hydroxyphenylpyruvic acid, phenylacetyl glycine and horse-urate, suggesting metabolic disorder of intestinal flora.Differentially expressed metabolites have a potential application value in the early diagnosis of zinc deficiency.

Serum Metabonomics Analysis of Diarrhea Mice Intervened by Zingiberis Rhizoma Recens Based on UPLC-QE-Orbitrap-MS / 中国实验方剂学杂志

ObjectiveTo investigate the antidiarrheal effect and mechanism of Zingiberis Rhizoma Recens on diarrhea mice， and to provide research basis for the inhibition of intestinal peristalsis by Zingiberis Rhizoma Recens and its application in the treatment of gastrointestinal diseases. MethodThe diarrhea model of mice was established by Sennae Folium. The control group， model group， Zingiberis Rhizoma Recens low-， medium-， high-dose groups （0.1， 0.32， 1.0 g·kg-1） and loperamide group （1.6 g·kg-1） were set. The intervention effect of Zingiberis Rhizoma Recens with different doses on diarrhea mice was detected by diarrhea score， incidence rate of loose stools （LSIR）， grade of average loose stools （ALSG）， diarrhea index （DI）， intestinal propulsion rate and intestinal pathological section. The serum metabonomics of mice was analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA）. The conditions were as follows：mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B）， flow rate of 0.4 mL·min-1， injection volume of 5 µL， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 100-1 500. ResultCompared with the model group， Zingiberis Rhizoma Recens high-dose group could obviously reduce the diarrhea score， LSIR， ALSG， DI and intestinal propulsion rate （P<0.05， P<0.01）， and improve the intestinal mucosal injury. There were 40 main differential metabolites among the control group， model group and Zingiberis Rhizoma Recens high-dose group， including glucose 1-phosphate， xanthine， xanthosine and so on. The metabolic pathways mainly included starch and sucrose metabolism， amino sugar and nucleotide sugar metabolism， fructose and mannose metabolism， tryptophan metabolism， and galactose metabolism. ConclusionZingiberis Rhizoma Recens can inhibit intestinal peristalsis in diarrhea mice and exert antidiarrhoea effect， the mechanism of which may be related to the regulation of carbohydrate and amino acid metabolism.

GC-MS Metabolites and Gene Expression Characteristics in Fibrous Roots of Dioscorea zingiberensis in Response to Low Phosphorus Stress / 中国实验方剂学杂志

ObjectiveTo investigate the metabolites and gene expression characteristics in fibrous roots of Dioscorea zingiberensis in response to low phosphorus stress. MethodThe severe stress group， the moderate stress group， and the normal group were set up to stimulate the low phosphorus stress experiment. The fibrous roots of D. zingiberensis were collected during initial stress. The metabolites and transcriptomic characteristics were analyzed by gas chromatography-mass spectrometry （GC-MS） derivatization and RNA-seq techniques. Through multivariate statistical analysis of metabolites treated by different methods，functional analysis of differentially expressed genes， and data mining， the metabolism markers produced in fibrous roots of D. zingiberensis under low phosphorus stress were screened out， and the metabolic pathway characteristics of different genes were analyzed. ResultA total of 116 GC-MS metabolites were detected from the fibrous roots of D. zingiberensis. The metabolic characteristics of fibrous roots of D. zingiberensis under different low phosphorus treatments were obviously different. Orthogonal partial least squares discriminant analysis（OPLS-DA） model was used to screen six differential metabolites represented by sugars and alcohols from metabolites of fibrous roots treated with different methods，and these components were presumedly metabolism markers of fibrous roots of D. zingiberensis in response to low phosphorus stress. The differential genes screened out from the severe stress group and the normal group were mainly enriched in peroxidase pathway，phosphate and hypophosphate metabolism pathway，while the differential genes screened out from the severe stress group and the moderate stress group were mainly enriched in glutathione metabolism pathway and phosphopentose pathway. A total of 177 differential genes in response to low phosphorus stress were screened out from fibrous roots， involving many pathways such as terpenoid skeleton and inositol biosynthesis，which was consistent with the fact that the metabolic differential components in fibrous roots in response to low phosphorus stress were mainly saccharides and inositol. ConclusionThe metabolites and gene expression in fibrous roots of D. zingiberensis responded to low phosphorus stress，and the differential metabolites were closely related to differentially expressed genes. This study is expected to provide a theoretical basis for the research on the molecular mechanism of D. zingiberensis in response to low phosphorus stress.

Metabonomic study of biochemical changes in serum of PCPA-induced insomnia rats after treatment with Suanzaoren Decoction / 中国中药杂志

Suanzaoren Decoction(SZRD) is a classical formula for the clinical treatment of insomnia. This study analyzed the effect of SZRD on endogenous metabolites in insomnia rats based on metabonomics and thereby explored the anti-insomnia mechanism of SZRD. To be specific, DL-4-chlorophenylalanine(PCPA) was used to induce insomnia in rats. Then pathological changes of the liver and brain were observed and biochemical indexes such as 5-hydroxytryptamine(5-HT), dopamine(DA), glutamate(Glu), γ-aminobutyric acid(GABA), and norepinephrine(NE) in the hippocampus and prostaglandin D2(PGD2), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and IL-6 in the serum of rats were detected. On this basis, the effect of SZRD on PCPA-induced insomnia rats was preliminarily assessed. The metabolic profile of rat serum samples was further analyzed by ultra-performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS). Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined with t-test and variable importance in projection(VIP) to identify differential metabolites, and MetaboAnalyst 5.0 was employed for pathway analysis. The results showed that SZRD could improve the pathological changes of brain and liver tissues, increase the levels of neurotransmitters 5-HT, DA, and GABA in hippocampus and the level of PGD2 in hypothalamic-pituitary-adrenal axis(HPA axis), and reduce the levels of IL-1β and TNF-α in serum of insomnia rats. Metabonomics analysis yielded 12 significantly changed potential metabolites: 5-aminovaleric acid, N-acetylvaline, L-proline, L-glutamate, L-valine, DL-norvaline, D(-)-arginine, pyroglutamic acid, 1-methylguanine, L-isoleucine, 7-ethoxy-4-methylcoumarin, and phthalic acid mono-2-ethylhexyl ester(MEHP), which were related with multiple biochemical processes including metabolism of D-glutamine and D-glutamate, metabolism of alanine, aspartate, and glutamate, metabolism of arginine and proline, arginine biosynthesis, glutathione metabolism. These metabolic changes indicated that SZRD can improve the metabolism in insomnia rats by regulating amino acid metabolism.

Effects of Angong Niuhuang Pills against heart failure based on cross-scale polypharmacological study / 中国中药杂志

Angong Niuhuang Pills(AGNHP) are effective in clearing heat, removing the toxin, and eliminating phlegm for resuscitation. Clinically, it is widely used to treat various diseases such as febrile convulsion due to heat attacking pericardium, but its therapeutic effects on heart failure(HF) have not been well recognized. In this study, the profiles of differential metabolites regulated by AGNHP were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The underlying mechanism of AGNHP against HF was illustrated based on the integrated analysis of pharmacological data and metabolic molecular network. The HF model was induced by isoproterenol in mice. After oral administration of AGNHP for one week, cardiac functions in HF mice were evaluated by echocardiography, and serum samples of mice were collected for metabolomics analysis. Eight differential metabolites of AGNHP against HF were screened out through partial least square discriminant analysis(PLS-DA) and input into MetaboAnalyst for the analysis of metabolic pathways. Moreover, the critical metabolic pathways regulated by AGNHP were enriched according to the potential targets of major compounds in AGNHP. After AGNHP treatment, the recovered index of relative content of some metabolites underwent cross-scale fusion analysis with therapeutic efficacy data, followed by "compound-reaction-enzyme-gene" network analysis. It is inferred that the anti-HF effects of AGNHP may be attributed to the metabolism of arachidonic acid, amino acid, glycerophospholipid, and linoleic acid. The cross-scale polypharmacological analysis method developed in this study provides a new method to interpret scientific principles of AGNHP against HF with modern technologies.

Metabolomics Analysis of Codonopsis pilosula Under Drought Stress Based on LC-MS / 中国实验方剂学杂志

Objective:The biological mechanism of <italic>Codonopsis pilosula</italic> adaptation to drought was explored by determining the root metabolome of <italic>C. pilosula</italic> during harvesting. Method:Non-targeted metabonomics LC-MS was used to screen differential metabolites by multivariate statistical analysis，univariate statistical analysis and metabolic pathway enrichment analysis. Result:①There were 274 metabolites in LD vs CK group，142 of which were up-regulated and 132 of which were down-regulated. There were 284 metabolites with significant difference in MD vs CK group，of which 157 were up-regulated and 127 were down-regulated. There were 317 metabolites with significant difference in SD vs CK group，of which 133 were up-regulated and 184 were down-regulated. ②Differential metabolites were annotated into kyoto Encyclopedia of Gene and Genomes （KEGG） database and 82 differential metabolic pathways were obtained，among which sphingolipids metabolism was significantly enriched （<italic>P</italic><0.01）.Metabolism of arginine and proline，tryptophan，alanine，galactose，nicotinic acid and nicotinamide，cysteine and methionine，arachidonic acid，linolenic acid and glycerides were significantly enriched in different metabolite pathways （<italic>P</italic><0.05）. ③The metabolites of the three comparison groups before and after enrichment were classified and analyzed. It was found that they were mainly concentrated in fatty acyls group，carboxylic acid and derivatives，and organ oxygen compounds，followed by sphingolipids，indoles and derivatives，organonitrogen compounds，glycerophospholipids，pyridines and derivatives，peptidomimetics，glycerolipids and so on.In the drought stress of <italic>C. codonopsis</italic>，carbohydrate related metabolites were mainly up-regulated，lipid related metabolites were mainly down-regulated，and all other metabolites were up-regulated. Conclusion:The changes of metabolites in the roots of <italic>C. pilosula</italic> under drought stress were elucidated. carbohydrate and lipid-related metabolites were the main products of <italic>C. pilosula</italic> under drought stress，and these metabolites may be the main reason to improve the ability of <italic>C. pilosula</italic> to resist drought，which laid a foundation for further study on the mechanism of <italic>C. pilosula</italic> to resist drought.

Serum Metabonomics Analysis of Colitis Associated Cancer Mice Intervened by Hedyotis diffusa Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:To investigate the intervention of <italic>Hedyotis diffusa</italic> （HDW） on colitis associated cancer （CAC） model mice and explore its mechanism. Method:The CAC mouse model was established by synergistic action of azoxymethane （AOM） and dextran sulfate sodium （DSS）. The intervention of HDW on CAC mice was evaluated by disease activity index （DAI）， colonic tissue morphology， pathological injury score and tumorigenesis rate. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS） and multivariate statistical analysis were used to analyze the metabonomics of mice serum and to explore the mechanism of HDW intervention on CAC. Result:HDW could significantly improve the general condition of CAC mice， decrease DAI， colon gross morphological score， histopathological score and tumorigenesis rate. Compared with the normal group， 38 kinds of differential metabolites were screened in the model group， including 11 potential biomarkers， involving 11 main metabolic pathways. HDW could significantly regulate 9 kinds of differential metabolites ［niacinamide， uridine， 4-pyridoxic acid， LysoPC （18∶0）， LysoPE （0∶0/20∶0）， myo-inositol， purine， sphinganine 1-phosphate and tetradecanedioic acid］ in the model group， including 2 kinds of potential biomarkers （myo-inositol and niacinamide）， and HDW could regulate nicotinate and nicotinamide metabolism and inositol phosphate metabolism. Conclusion:HDW has a therapeutic effect on CAC， which may be achieved by regulation of energy metabolism and glucose metabolism.

Effect of Danhong Injection on platelet metabolites based on metabonomics technology / 中国中药杂志

The effect of Danhong Injection on the endogenous metabolites of rabbit platelets was analyzed by the liquid chromatography-mass spectrometry( LC-MS) based metabonomic approach. Anti-platelet aggregation was detected after Danhong Injection treatment and the changes of platelet metabolites were analyzed by metabonomics. Principal component analysis( PCA) and partial least squares discriminant analysis( PLS-DA) were performed to investigate the effect of Danhong Injection on endogenous metabolites of platelets,characterize the biomarkers,and explore the relevant pathways and the underlying mechanism. As demonstrated by the pharmacodynamic results,Danhong Injection of different doses and concentrations antagonized platelet aggregation in a dose-and concentration-dependent manner. In contrast to the control group,25 differential metabolites such as nicotinic acid,nicotinic acid riboside,and hypoxanthine were screened out after platelets were treated by Danhong Injection. These metabolites,serving as important biomarkers,were mainly enriched in the nicotinic acid-niacinamide metabolic pathway and purine metabolic pathway. This study explored the therapeutic mechanism of Danhong Injection from a microscopic perspective by metabonomics,which is expected to provide a new idea for the investigation of platelet-related mechanisms.

Effect of light at night on liver metabolism in miceby untargeted metabolomics / 预防医学

Objective@#To observe the changes of liver metabolism in mice exposed to artificial light at night. @*Methods@#Healthy male C57BL/6J mice were randomly divided into the light at night group and the control group, with 8 mice in each group. The daily light/dark cycle was 12/12 hours in the control group, and 24/0 hours in the light at night group for 10 consecutive days. The hepatic metabolite profiles of the two groups of mice were detected by high performance liquid chromatography tandem mass spectrometry. The modelling was assessed by combining principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis ( OPLS-DA ) , The changes of metabolites in the two groups were compared through KEGG database.@*Results@#Compared with the control group, 9 different metabolites were detected in the light at night group, among which the down-regulated metabolites were glycine-betaine, glutathione, tyrosine, betaine, lysine, hypoxanthine, histidine and methionine, and the up-regulated ones were mannose-6-phosphate. The weight analysis of the metabolic pathways showed that the major influences on liver of light at night group were phenylalanine-tyrosine-tryptophan metabolism, tyrosine metabolism, fructose and mannose metabolism, cysteine and methionine metabolism and histidine metabolism. @*Conclusion@#The metabolism of various amino acids and sugars in light at night mice is disturbed,and the key differential metabolites are tyrosine, methionine, histidine and mannose-6-phosphate.

Metabonomics study of Shouhui Tongbian Capsules in slow transit constipation based on UPLC-ESI-QE-Orbitrap-MS / 中国中药杂志

The effect of Shouhui Tongbian Capsules(SHTB) on the endogenous metabolites of colon tissue in mice with slow transit constipation was analyzed by metabolomics methods to explore its mechanism in the treatment of constipation. ICR mice were randomly divided into normal group, model group and SHTB group according to the body weight. The mice were given diphenoxylate to establish the slow transit constipation model. Mouse carbon ink pushing rate, first defecation time and the number of defecation particles in 12 h were observed. The mouse colon tissue was separated and the mucous cells were detected by Periodic acid Schiff and Alcian blue(AB-PAS) staining. Ultra-high-performance liquid chromatography electrospray ionization orbitrap tandem mass spectrometry(UPLC-ESI-Orbitrap-MS/MS) technology was used to characterize the differences in tissue metabolism to screen out the potential different metabolites and possible metabolic pathways in colon tissue. The results indicated that SHTB could significantly shorten the first defecation time and the number of defecations, and increase the number of intestinal peristalsis and mucous cells in the colonic mucosa compared to the model mice. Metabolomics results showed that, compared with the normal group, a total of 17 potential biomarkers, including L-kynurenine, N6,N6,N6-trimethyl-L-lysine, L-formylkynurenine, N6-acetyl-L-lysine, L-phenylalanine, phenylacetaldehyde, xanthoxin, thymidine, glycyl-L-leucine, cystathionine,(R)-1-aminopropan-2-ol, deoxycytidine, gamma-glutamyl-gamma-aminobutyraldehyde, D-galactose, L-arginine, L-proline and pyruvate, were found and identified in colon tissue. Treated with SHTB, these metabolic differences tended to return to normal levels. Therefore, it could be made a conclusion that the therapeutic effect of SHTB on chronic transit constipation may be related to regulating phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arginine and proline metabolism, cysteine and methionine metabolism, tyrosine metabolism, arginine biosynthesis, pyruvate metabolism, glycolysis, pyrimidine metabolism, tricarboxylic acid cycle and galactose metabolism.

Abnormal Circadian Clock Ｐromotes AOM / DSS-induced Colon Carcinogenesis in Mice by Remodeling Tumor Metabolism / 中国生物化学与分子生物学报

Abnormal circadian clock has been identified as an independent risk factor for tumorigenesis, and is closely related to the occurrence and development of tumor. As metabolic disorder is also one of the important characteristics of tumorigenesis, therefore it is particularly important to investigate the regulatory relationship between biological clock and tumor metabolism. In this study, the effect of abnormal circadian clock on colon cancer growth was evaluated by azoxymethane (AOM) / dextran sodium sulfate (DSS) -induced colitis-associated carcinogenesis (CAC) mice model. The result showed that abnormal circadian clock aggravated anal swelling, redness, bloody and anorectal prolapse in CAC mice, and significantly increased the number and volume of CAC polyps (P <0. 05 or P <0. 01), and reduced the intestinal length, body weight, survival rate of CAC mice and the expression levels of inflammatory factors IL-1β (interleukin-1 beta) and TNFα (tumor necrosis factor α) (P < 0. 05 or P < 0. 01), indicating that abnormal biological clock promotes the occurrence and development of CAC. Further, non-target metabonomics analysis of serum samples from mice was performed by liquid chromatography-mass spectrometry (LC-MS) . The result showed that compared with CAC mice with normal circadian rhythm, 27 differential metabolites were identified in CAC mice with disrupted circadian clock, and 9 metabolic pathways were enriched by KEGG (kyoto encyclopedia of genes and genomes) database. These results suggest that abnormal circadian clock can significantly change the relative abundance of some metabolites in serum samples from CAC mice, remodel tumor metabolism, and result in the development of CAC in mice. This study reveals the pivotal role of tumor metabolism in the abnormal circadian clock promoting the growth of CAC in mice, providing a new experimental basis for the interaction between circadian clock and metabolic homeostasis in the occurrence and development of colon cancer.

Study on intervention of triptolide on metabolism of LC/IT-TOF-MS in ulcerative colitis / 中草药

Objective: The effect of triptolide (TP) on endogenous metabolites in mice with ulcerative colitis (UC) was analyzed by means of metabolomics, and the metabolic pathway and possible mechanism of TP in UC were discussed. Methods: C57BL/6 mice were randomly divided into blank control group, model group, and triptolide group. Dextran sulfate (DSS) was used to induce UC mice model. The serum samples of mice were detected by high performance liquid chromatography-mass spectrometry and characterized by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to identify the potential biomarkers and possible metabolic pathways. Results: Compared with the blank control group, a total of 15 potential biomarkers, such as cholic acid, bezoar cholic acid, goose-deoxycholic acid, citrulline, guanidine butyric acid, aminoacetic acid, and cis-aconitic acid, were found and identified in serum. Compared with the model group, the potential biomarkers showed a tendency of callback to normal level after TP intervention. Conclusion: Metabolomics analysis reveals that TP had certain therapeutic effects on UC mice, and its mechanism may be related to regulating primary bile acid biosynthesis, arginine, and proline metabolism.