Endogenous Metabolites In Serum Research Articles

Serum and Urine Metabolites and Kidney Function.

Key Points We provide an atlas of cross-sectional and longitudinal serum and urine metabolite associations with eGFR and urine albumin-creatinine ratio in an older community-based cohort.Metabolic profiling in serum and urine provides distinct and complementary insights into disease. Background Metabolites represent a read-out of cellular processes underlying states of health and disease. Methods We evaluated cross-sectional and longitudinal associations between 1255 serum and 1398 urine known and unknown (denoted with “X” in name) metabolites (Metabolon HD4, 721 detected in both biofluids) and kidney function in 1612 participants of the Atherosclerosis Risk in Communities study. All analyses were adjusted for clinical and demographic covariates, including for baseline eGFR and urine albumin-creatinine ratio (UACR) in longitudinal analyses. Results At visit 5 of the Atherosclerosis Risk in Communities study, the mean age of participants was 76 years (SD 6); 56% were women, mean eGFR was 62 ml/min per 1.73 m2 (SD 20), and median UACR level was 13 mg/g (interquartile range, 25). In cross-sectional analysis, 675 serum and 542 urine metabolites were associated with eGFR (Bonferroni-corrected P < 4.0E-5 for serum analyses and P < 3.6E-5 for urine analyses), including 248 metabolites shared across biofluids. Fewer metabolites (75 serum and 91 urine metabolites, including seven metabolites shared across biofluids) were cross-sectionally associated with albuminuria. Guanidinosuccinate; N2,N2-dimethylguanosine; hydroxy-N6,N6,N6-trimethyllysine; X-13844; and X-25422 were significantly associated with both eGFR and albuminuria. Over a mean follow-up of 6.6 years, serum mannose (hazard ratio [HR], 2.3 [1.6–3.2], P = 2.7E-5) and urine X-12117 (HR, 1.7 [1.3–2.2], P = 1.9E-5) were risk factors of UACR doubling, whereas urine sebacate (HR, 0.86 [0.80–0.92], P = 1.9E-5) was inversely associated. Compared with clinical characteristics alone, including the top five endogenous metabolites in serum and urine associated with longitudinal outcomes improved the outcome prediction (area under the receiver operating characteristic curves for eGFR decline: clinical model=0.79, clinical+metabolites model=0.87, P = 8.1E-6; for UACR doubling: clinical model=0.66, clinical+metabolites model=0.73, P = 2.9E-5). Conclusions Metabolomic profiling in different biofluids provided distinct and potentially complementary insights into the biology and prognosis of kidney diseases.

Bletilla striata polysaccharide improves toxic and side effects induced by 5-FU: an untargeted metabolomics study

This study aimed to analyze the effect of Bletilla striata polysaccharide(BSP) on endogenous metabolites in serum of tumor-bearing mice treated with 5-fluorouracil(5-FU) by untargeted metabolomics techniques and explore the mechanism of BSP in alleviating the toxic and side effects induced by 5-FU. Male BALB/C mice were randomly divided into a normal group, a model group, a 5-FU group, and a 5-FU + BSP group, with eight mice in each group. Mouse colon cancer cells(CT26) were transplanted into the mice except for those in the normal group to construct the tumor-bearing mouse model by subcutaneous injection, and 5-FU chemotherapy and BSP treatment were carried out from the second day of modeling. The changes in body weight, diarrhea, and white blood cell count in the peripheral blood were recorded. The mice were sacrificed and sampled when the tumor weight of mice in the model group reached approximately 1 g. TUNEL staining was used to detect the cell apoptosis in the small intestine of each group. The proportions of hematopoietic stem cells and myeloid progenitor cells in bone marrow were measured by flow cytometry. Five serum samples were selected randomly from each group for untargeted metabolomics analysis. The results showed that BSP was not effective in inhibiting colon cancer in mice, but diarrhea, leukopenia, and weight loss caused by 5-FU chemotherapy were significantly improved after BSP intervention. In addition, apoptotic cells decreased in the small intestinal tissues and the percentages of hematopoietic stem cells and myeloid progenitor cells in bone marrow were significantly higher after BSP treatment. Metabolomics results showed that the toxic and side effects of 5-FU resulted in significant decrease in 29 metabolites and significant increase in 22 metabolites in mouse serum. Among them, 19 disordered metabolites showed a return to normal levels in the 5-FU+BSP group. The results of pathway enrichment indicated that metabolic pathways mainly involved pyrimidine metabolism, arachidonic acid metabolism, and steroid hormone biosynthesis. Therefore, BSP may ameliorate the toxic and side effects of 5-FU in the intestinal tract and bone marrow presumably by regulating nucleotide synthesis, inflammatory damage, and hormone production.

Metabonomics comparison of coronary heart disease with and without turbid phlegm syndrome

Objectives To help understand the pathogenesis of coronary heart disease (CHD) by doing blood metabonomics comparison on CHD patients with phlegm-turbidity syndrome and without syndrome, and healthy controls. Methods Ninety patients were involved. Among them, 60 patients with CHD were divided into the coronary heart disease with turbid phlegm syndrome group (HZTZ) and the coronary heart disease without turbid phlegm syndrome group (NHZTZ). The additional 30 subjects were the healthy controls (JKZ). The serum endogenous metabolites were determined and analyzed by Liquid chromatography—Mass spectrometry (LC-MS). Results In positive ion mode, there were three identical differential metabolites among the three groups: N2-ethyl-n4-isopropyl-6-(methylthio)-1,3,5-triazine-2,4-diamine, LysoPC (18:2), and methyl palmitate. In negative ion mode, two identical metabolites were identified, which were LysoPC (17:0) and fatty acyl ester of hydroxy fatty acids (FAHFA) (17:1/14:1). These five metabolites of the HZTZ group were higher than those in NHZTZ group and healthy controls. The results identified perturbations in the areas, such as phenylalanine, tyrosine, and tryptophan biosynthesis, starch and sucrose metabolism, carbohydrate digestion and absorption, and so on. Conclusions Metabonomics analysis of serum in patients with CHD with turbid phlegm syndrome uncovered the metabolic disorders of saturated and unsaturated fatty acids in comparison to the patients without turbid phlegm syndrome.

UPLC-Q-Exactive/MS-based metabonomics revealed protective effect of Zingiberis rhizome and its processed product on deficiency-cold and hemorrhagic syndrome rats.

Zingiberis rhizome carbonisata (ZRC) is the processed product of Zingiberis rhizome (ZR). ZR is mainly used for warming the spleen and stomach to dispel cold, whereas ZRC is commonly applied as a treatment for deficiency-cold and hemorrhagic syndrome (DCHS). Although they have long been used to serve different clinical purposes, the specific action mechanism of the drugs and molecular changes underlying ZR processing are not clear. In this study, metabolomics study was carried out to analyze the alterations in endogenous metabolites in serum and urine samples of DCHS rat models using ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry technique and constructed principal component analysis score plots that showed that the ZRC group was completely separated from the DCHS and ZR groups but demonstrated a highly close plotting to the normal control group. The results revealed that both ZR and ZRC intervened in the metabolic pathways of DCHS models but to varying degrees and with different influencing factors. In addition, ZRC was found to function as a treatment for the metabolic disorders of DCHS through 15 pharmacodynamic biomarkers involving a series of pathways, such as glycine, serine, and threonine metabolic pathway, as well as arachidonic acid metabolic pathways. This study showed that metabolomics method based on ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry could preliminarily illuminate the therapeutic mechanism of ZR and ZRC on DCHS and the changes in ZR processing from the molecular-level perspective. The results also provided new insight into further research on DCHS treatment.

Metabolomics analysis reveals the renal protective effect of Panax ginseng C. A. Mey in type 1 diabetic rats.

The dry root and rhizome of Panax ginseng C. A. Mey has garnered much interest owing to its medicinal properties against diabetes and cardiovascular diseases. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS)-based metabolomics approach was used to illustrate the therapeutic mechanisms of ginseng extract on the serum and urinary metabolic profiles in streptozotocin-induced type 1 diabetes mellitus (T1DM) rats. Pharmacological and renal parameters in response to the administration of ginseng were also evaluated. In total, 16 serum endogenous metabolites and 14 urine endogenous metabolites, including pyruvic acid, indoleacetic acid, and phenylacetylglycine, were identified as potential biomarkers for diabetes. Pathway enrichment and network analysis revealed that the biomarkers modulated by ginseng were primarily involved in phenylalanine and pyruvate metabolism, as well as in arginine biosynthesis. Moreover, the levels of several renal injury-related biomarkers in T1DM rats were significantly restored following treatment with ginseng. The administration of the extract helped maintain tissue structure integrity and ameliorated renal injury. The findings suggest that the regulatory effect of ginseng extract on T1DM involves metabolic management of diabetic rats, which subsequently attenuates T1DM-induced early renal dysfunction.

Combined metabolomics and machine learning algorithms to explore metabolic biomarkers for diagnosis of acute myocardial ischemia.

Acute myocardial ischemia (AMI) remains the leading cause of death worldwide, and the post-mortem diagnosis of AMI represents a current challenge for both clinical and forensic pathologists. In the present study, the untargeted metabolomics based on ultra-performance liquid chromatography combined with high-resolution mass spectrometry was applied to analyze serum metabolic signatures from AMI in a rat model (n = 10 per group). A total of 28 endogenous metabolites in serum were significantly altered in AMI group relative to control and sham groups. A set of machine learning algorithms, namely gradient tree boosting (GTB), support vector machine (SVM), random forest (RF), logistic regression (LR), and multilayer perceptron (MLP) models, was used to screen the more valuable metabolites from 28 metabolites to optimize the biomarker panel. The results showed that classification accuracy and performance of MLP model were better than other algorithms when the metabolites consisting of L-threonic acid, N-acetyl-L-cysteine, CMPF, glycocholic acid, L-tyrosine, cholic acid, and glycoursodeoxycholic acid. Finally, 17 blood samples from autopsy cases were applied to validate the classification model's value in human samples. The MLP model constructed based on rat dataset achieved accuracy of 88.23%, and ROC of 0.89 for predicting AMI type II in autopsy cases of sudden cardiac death. The results demonstrated that MLP model based on 7 molecular biomarkers had a good diagnostic performance for both AMI rats and autopsy-based blood samples. Thus, the combination of metabolomics and machine learning algorithms provides a novel strategy for AMI diagnosis.

Serum metabolomic responses to aerobic exercise in rats under chronic unpredictable mild stress

This study analyzed the effects of aerobic exercise on endogenous serum metabolites in response to chronic unpredictable mild stress (CUMS) using a rat model, aiming to identify the metabolic regulatory pathways involved in the antidepressant effect resulted from a 28-day treadmill aerobic exercise intervention. The animals were randomly divided into four groups (n = 8): normal control, normal with aerobic exercise, CUMS control, and CUMS with aerobic exercise. Body weight, sucrose preference and open field tests were performed weekly during the intervention period for changes in depressant symptoms. Serum metabolic profiles obtained by using the LC–MS/MS metabolomics were analyzed to explore the regulatory mechanism for the effect of the aerobic exercise on depression. Behavior tests showed that the aerobic exercise resulted in a significant improvement in depression-like behavior in the CUMS rats. A total of 21 differential metabolites were identified as being associated with depression in serum metabolic profile, of which the aerobic exercise significantly modulated 15, mainly related to amino acid metabolism and energy metabolism. Collectively, this is the first study that LC–MS/MS techniques were used to reveal the modulatory effects of aerobic exercise on the serum metabolic profile of depressed rats and the findings further enriched our understanding of potential mechanisms of aerobic exercise interventions on depression.

UHPLC-MS-Based Serum and Urine Metabolomics Reveals the Anti-Diabetic Mechanism of Ginsenoside Re in Type 2 Diabetic Rats.

Panax ginseng was employed in the treatment of “Xiao-Ke” symptom, which nowadays known as diabetes mellitus, in traditional Chinese medicine for more than a thousand years. Ginsenoside Re was the major pharmacologic ingredient found abundantly in ginseng. However, the anti-diabetic of Ginsenoside Re and its underlying mechanism in metabolic level are still unclear. Serum and urine metabolomic method was carried out to investigate the anti-diabetic pharmacological effects and the potential mechanism of Ginsenoside Re on high-fat diet combined streptozotocin-induced type 2 diabetes mellitus (T2DM) rats based on ultra-high-performance liquid chromatography coupled with quadrupole exactive orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap/MS). Serum and urine samples were collected from the control group (CON), T2DM group, metformin (MET) treatment group, and ginsenoside Re treatment group after intervention. The biochemical parameters of serum were firstly analyzed. The endogenous metabolites in serum and urine were detected by UHPLC-MS. The potential metabolites were screened by multivariate statistical analysis and identified by accurate mass measurement, MS/MS, and metabolite databases. The anti-diabetic-related metabolites were analyzed by KEGG metabolic pathway, and its potential mechanism was discussed. The treatment of ginsenoside Re significantly reduced the blood glucose and serum lipid level improved the oxidative stress caused by T2DM. Biochemical parameters (urea nitrogen, uric acid) showed that ginsenoside Re could improve renal function in T2DM rats. Respective 2 and 6 differential metabolites were found and identified in serum and urine of ginsenoside Re compared with T2DM group and enriched in KEGG pathway. Metabolic pathways analysis indicated that the differential metabolites related to T2DM were mainly involved in arachidonic acid metabolism, Vitamin B6, steroid hormone biosynthesis, and bile secretion metabolic pathways. This study verified the anti-diabetic and anti-oxidation effects of ginsenoside Re, elaborated that ginsenoside Re has a good regulation of the metabolic disorder in T2DM rats, which could promote insulin secretion, stimulated cannabinoid type 1 receptor (CB1), and CaMKK β to activate AMPK signaling pathway, inhibited insulin resistance, and improved blood glucose uptake and diabetic nephropathy, so as to play the role of anti-diabetic.

Analgesic Activity of Jin Ling Zi Powder and Its Single Herbs: A Serum Metabonomics Study.

To compare the analgesic effect of Jin Ling Zi Powder (JLZ) and its two single herbs. The hot plate method was used to induce pain. Totally 36 mice were randomly divided into 6 groups by a complete random design, including control, model, aspirin (ASP, 0.14 g/kg body weight), JLZ (14 g/kg body weight), Corydalis yanhusuo (YHS, 14 g/kg body weight), and Toosendan Fructus (TF, 14 g/kg body weight) groups, 6 mice in each group. The mice in the control and model groups were given the same volume of saline, daily for 2 consecutive weeks. At 30, 60, 90, and 120 min after the last administration, the pain threshold of mice in each group was measured, and the improvement rate of pain threshold was calculated. Serum endogenous metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS). There was no statistical difference in pain threshold among groups before administration (P>0.05). After 2 weeks of administration, compared with the model group, the pain threshold in JLZ, YHS, TF and ASP groups were increased to varying degrees (P<0.05). JLZ had the best analgesic effect and was superior to YHS and TF groups. A total of 14 potential biomarkers were screened in serum data analysis and potential biomarkers levels were all reversed to different degrees after the treatment with JLZ and its single herbs. These potential biomarkers were mainly related to glyoxylate and dicarboxylate metabolism, glycine, serine and threonine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and inositol phosphate metabolism. The analgesic mechanism of JLZ and YHS was mainly due to the combination of glycine and its receptor, producing post-synaptic potential, reducing the excitability of neurons, and weakening the afferent effect of painful information.

Chemical Fingerprint Analysis and Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry-Based Metabolomics Study of the Protective Effect of Buxue Yimu Granule in Medical-Induced Incomplete Abortion Rats.

Medical abortion is a common method to terminate an early pregnancy and often causes serious complications such as abnormal uterine bleeding and endometritis. Buxue Yimu granule (BYG) is a well-known traditional Chinese medicine prescription composed of five kinds of drugs and is widely used in gynecology and obstetrics. The aim of the present study was to establish the quality standard of BYG and investigate its protective effect on incomplete abortion. The chemical fingerprint of BYG was established by high performance liquid chromatography (HPLC). The major compounds of BYG were determined by ultra-performance liquid chromatography with triple quadrupole mass spectrometry. An incomplete abortion rat model was induced by intragastric administration of mifepristone (8.3 mg·kg−1) combined with misoprostol (100.0 μg·kg−1) during early pregnancy. The serum levels of human chorionic gonadotrophin (HCG), estradiol (E2), and progesterone (PG) were determined. The serum endogenous metabolites were analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Multivariate analysis, including partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA), was employed to analyze the metabolic profiles, and MetaboAnalyst was used to investigate the metabolic pathways. Furthermore, hematoxylin-eosin staining (HE) was used to evaluate the histopathological changes in uterine tissue. The expression levels of VEGFA and NF-κB were detected by immunohistochemistry. The results indicated that HPLC fingerprint analysis can be successfully used to assess the quality of BYG. The medical-induced incomplete abortion rats were clearly separated from control rats, and the biochemical changes were gradually restored to normal after administration of BYG. Moreover, 19 potential biomarkers, including N-lactoylleucine, 2-piperidinone, isobutyryl-l-carnitine, eicosapentaenoylcholine, LysoPC(14:0), LysoPC(20:5), physagulin C, LysoPC(18:3), leukotriene D5, deoxycholic acid 3-glucuronide, glycine, pregnanediol 3-O-glucuronide, LysoPC(18:2), LysoPC(17:0/0:0), N-acetyl-leukotriene E4, LysoPC(18:0), platelet-activating factor, LysoPA(24:1), and LysoPC(18:1), which were mainly related to the amino acids metabolism, lipids metabolism, and bile acid biosynthesis, were identified. Consequently, BYG exerts a potential protective role in the intervention of incomplete abortion by anti-inflammatory, promote endometrial repair, and regulate the metabolic disorders.

Metabonomics study on orthotopic transplantion mice model of colon cancer treated with Astragalus membranaceus-Curcuma wenyujin in different proportions via UPLC-Q-TOF/MS

Metabolomics, an important part of systems biology, can reveal the complex pathogenesis of many diseases and mechanism of Chinese materia medica (CMM). Astragalus membranaceus-Curcuma wenyujin (AC) was a classic drug pair that has a good clinical effect on gastrointestinal inflammation and many tumors. Our previous research proved that AC can inhibit tumor growth and metastasis especially the colorectal cancer (CRC), also promote the normalization of tumor blood vessels, but its optimal ratio and the specific mechanism is still not clear. In this study, colon cancer mice of orthotopic transplantion model was used to screen the best proportion, UPLC-Q-TOF/MS metabolomics analysis method was established to explore the pathogenesis of colon cancer and the molecular mechanism of AC. The correlation analysis of metabolite changes and tumor growth was analyzed by R language. The result showed that AC at the ratio of 2:1 showed the best effect on inhibiting tumor growth, also the liver and spleen metastasis rate. A total of 23 potential biomarkers were detected in the serum of colon cancer mice by the analysis of Progenesis QI (Version 2.4) software. Among this, 11 metabolites including purines, steroids, phytosphingosine and l-palmitoylcarnitine were up-regulated in CC mice, while 12 metabolites like amino acids, deoxyribose and dihydrobiopterin were down-regulated in CC mice. After the treatment of AC for 15 days, 8 biomarkers were up-regulated, and 9 biomarkers down-regulated. Especially, AC at the ratio of 2:1 showed a significant callback effect on metabolic biomarkers, such as hypoxanthine, xanthosine, 7-methylxanthine, all-trans-retinoic acid, dihomo-γ-linolenic acid. 8 metabolic pathways: Aminoacyl-tRNA biosynthesis, Nicotinate and nicotinamide metabolism, Phenylalanine, tyrosine and tryptophan biosynthesis, Valine, leucine and isoleucine biosynthesis, Phenylalanine metabolism, Caffeine metabolism, Retinol metabolism, Alanine, aspartate and glutamate metabolism were selected as the model group disturbed metabolic pathways after the enrichment of MetaboAnalyst 4.0 online analysis software. And compared with the model group, Valine, leucine and isoleucine biosynthesis, Aminoacyl-tRNA biosynthesis, Caffeine metabolism pathway and Retinol metabolism pathways were altered after the intervention of AC. The correlation analysis results showed that various endogenous metabolites in serum have a strong correlation with tumor weight, such as hypoxanthine, which provides a basis for the selection of clinical markers. The results showed that AC can partially regulate metabolic disorder of CC mice by reversing the changes of metabolites, so as to inhibit the growth and metastasis of colon cancer, especially at the ratio of 2:1. These findings can provide a scientific basis for exploring the diagnostic biomarkers of colon cancer, and for clinical application of AC in the treatment of CRC program.

Attribution and identification of absorbed components by HPLC-DAD-ESI-MS after oral administration of Erhuang decoction

To realize the attribution and identification of absorbed components in rat serum after oral administration of Erhuang decoction prepared by semi-bionic enzyme extraction method, the fingerprints of serum samples were established using a HPLC-DAD-ESI-MS method. Thirty-two peaks in Erhuang decoction and 24 peaks in rat serum after oral administration of Erhuang decoction were detected. Among the 24 peaks detected in rat serum, 25 compounds were identified by comparing the retention time and mass spectrometry data with that of reference compounds, or by mass spectrometry analysis and retrieving the reference literatures. Among the identified 25 compounds in vivo, 24 were the original form of compounds absorbed from the detected compounds in vitro, and one was the metabolite compounds of licorice. By analyzing the mass spectrometry or ultraviolet absorption characteristics, other unidentified compounds in vivo were deduced to be the endogenous metabolites in serum or the original form and metabolites of the compounds existed in vivo. Results indicated that HPLC-DAD-ESI-MS is suitable for identifying the bioactive constituents in serum after oral administration of Erhuang decoction, and the findings would be beneficial to further research and development of the pharmacodynamic substance base of Erhuang decoction.

Anti-inflammatory mechanism of Crepis crocea based on NF-κB signaling pathway and ~1H-NMR metabonomics

Based on ~1H-NMR metabonomics technique and Western blot assay, the anti-inflammatory mechanism of Crepis crocea was discussed. In this study, male SD rats were treated with water extract(2.5 g·kg~(-1)) and dexamethasone acetate(6.25×10~(-4) g·kg~(-1)) for one week, and the inflammation model was induced by lipopolysaccharide(LPS). Then the counts of inflammatory cells white blood ceel(WBC), eosinophil(EO), lymphocyte(LY), basophils(BA) and neutrophils(NE) in whole blood of rats were observed. The levels of serum inflammatory factors tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6 and the expression of nuclear factor-κB(NF-κB) signaling pathway p65 and p-IκBα proteins in lung tissues were detected, and the change rules of serum endogenous metabolites were analyzed by ~1H-NMR metabonomics technique. The levels of TNF-α, IL-1β, IL-6 and NF-κB signaling pathway p65 and p-IκBα proteins were combined with ~1H-NMR metabonomics to study the anti-inflammatory mechanism of C. crocea. The results showed that the water extract of C. crocea significantly decreased the number of WBC, NE, EO, increased the number of BA and LY, decreased the levels of TNF-α, IL-1β, IL-6 and the expression of p65 and p-IκBα protein in NF-κB signaling pathway, and effectively alleviated the inflammatory symptoms. In the correlation analysis of differential metabolites regulated of C. crocea, four significant metabolites were obtained, including glycine, creatine, methionine and succinic acid. The anti-inflammatory mechanism of C. crocea may be related to the decrease of TNF-α, IL-1β, IL-6 levels and the protein expression of NF-κB signaling pathway, as well as the regulation of glycine, creatine, methionine and succinic acid metabolism.

Mass spectrometry-based metabolomics identifies the effects of dietary oligosaccharide-zinc complex on serum and liver of zinc deficiency mice

Abstract The present study aimed to investigate the metabolic mechanisms of zinc-deficient mice and oligosaccharide-zinc diets intervention by assessing the changes of endogenous metabolites in serum and liver using LC-MS metabolomics approach. Three (3) to four (4) weeks old Kunming mice were divided into three groups: control (normal diet), zinc-deficiency (without zinc diet), oligosaccharide-zinc (without zinc diet + oligosaccharide-zinc at a dose of 3.48 mg Zn/kg·bw). After 16 days treatment of diet with oligosaccharide-zinc complex, metabolites, such as lipid metabolites (fatty acids, phosphatidylcholines (PCs), lysophosphatidylcholines (lysoPCs), lipid metabolism intermediates (betaine), glycerophospholipid, phospholipid, dihydroxyacetone, pyrimidine and aldonic acid were significantly affected. The PLS-DA score plots showed a clear distinction of metabolites between experimental groups in serum and liver markers. Oligosaccharide-zinc treatment recovered the major metabolites changes in serum and liver in zinc-deficiency mice. Those results suggested that oligosaccharide-zinc complex could be regarded as a favourable zinc supplementation.

Metabolic effect of AOS-iron in rats with iron deficiency anemia using LC-MS/MS based metabolomics.

Iron deficiency anemia (IDA) is a worldwide nutritional problem. The metabolic mechanism of IDA is still unclear. So, the underlying metabolic mechanism of iron supplementation has not been reported even if various iron supplements to treat IDA have been studied. The present study aimed to investigate the metabolic mechanisms of IDA and agar oligosaccharide-iron complex (AOS-iron) supplementation in IDA rats by assessing the changes of endogenous metabolites in serum and liver using LC-MS/MS metabolomics approach. Orthogonal partial least-squares discriminant analysis (OPLS-DA) score plots showed significant separation of metabolites in serum and liver among the normal, anemia model and AOS-iron groups. Seventeen and eight metabolites were identified from serum and liver, respectively. Pathway enrichment analysis suggested that potential biomarkers were strongly involved in the biosynthesis of saturated and unsaturated fatty acids, sphingolipid metabolism, glycerophospholipid metabolism, linoleic acid metabolism, Fcγ receptor (FcγR)-mediated phagocytosis, pancreatic cancer metabolism, regulation of autophagy, gonadotropin releasing hormone (GnRH) signaling pathway, fatty acid metabolism, pantothenate and CoA biosynthesis, glutathione metabolism and primary bile acid biosynthesis. After supplementing 2mg Fe/kg·bw AOS-iron for 4weeks, the major metabolites in related pathways disrupted by IDA were restored to normal levels. Therefore, AOS-iron effectively treated IDA by regulating metabolic disorders.

Serum and urine metabolomics based on UPLC-Q-TOF/MS reveals the antipyretic mechanism of Reduning injection in a rat model.

Reduning injection (RDN), a patented traditional Chinese medicine, has the obvious antipyretic effect and has been widely used in China. Although some previous studies proved its antipyretic effect by animal efficacy experiment or clinical observation, its holistic mechanism in vivo was still unclear. To comprehensively elucidate the antipyretic mechanism of RDN, the investigation of fever-related potential biomarkers and metabolic pathways in the rat fever model is described in this paper. Rat fever model was established by dry yeast. A large number of endogenous metabolites in serum and urine were detected by UPLC-Q-TOF/MS, and fever-related potential biomarkers were screened and identified by multivariate analysis and metabolite databases. The reliability and biological significance of the largely disturbed biomarkers was verified by the metabolic network and the correlation with pharmacodynamic indicators, which contained IL-1β, IL-6, TNF-α, PGE2 and cAMP. The established UPLC-Q-TOF/MS analytical method afforded satisfactory results in terms of precision, repeatability and stability, which met the requirements of biological sample determination. A total of 32 potential biomarkers associated with fever were screened and identified, among which 22 species could be adjusted by RDN. The metabolism pathway analysis revealed that valine, leucine and isoleucine biosynthesis, and sphingolipid metabolism were greatly disturbed. Their biomarkers involved L-leucine, L-valine, sphinganine and phytosphingosine, all of which showed a callback trend after RDN was given. These 4 biomarkers had a certain correlation with some known fever-related small molecules and pharmacodynamic indicators, which indicated that the selected fever-related biomarkers had certain reliability and biological significance. RDN has a good regulation of the metabolic disorder of endogenous components in dry yeast-induced fever rats.Its antipyretic mechanism is mainly related to the regulation of amino acid, lipid and energy metabolism. The study is useful to better understand and analyze the pharmacodynamic mechanism of complex systems, such as traditional Chinese medicine.

Dendrobium officinale Polysaccharides Inhibit 1-Methyl-2-Nitro-1-Nitrosoguanidine Induced Precancerous Lesions of Gastric Cancer in Rats through Regulating Wnt/β-Catenin Pathway and Altering Serum Endogenous Metabolites.

Dendrobium officinale is a herb in traditional Chinese medicine where D. officinale polysaccharides (DOP) are the main active ingredient. This study aimed at evaluating DOP efficiency at inhibiting 1-Methyl-2-nitro-1-nitrosoguanidine (MNNG) induced precancerous lesions of gastric cancer (PLGC) in rats through the Wnt/b-catenin pathway and analyzing the variations of serum endogenous metabolites. PLGC was established in male Sprague-Dawley (SD) rats by administering 150 μg/mL MNNG in drinking water for 7 months and giving 0.1 mL of 10% NaCl once weekly during the initial 20 weeks. Treatment with DOP inhibited the progress of PLGC through decreasing the expression of β-catenin by immunohistochemical analysis. The futher study indicated DOP downregulated gene expression of Wnt2β, Gsk3β, PCNA, CyclinD1, and β-catenin, as well as protein expression of Wnt2β, PCNA, and β-catenin. On the other hand, there were nine endogenous metabolites identified after the DOP treatment. Among these, the most significant one is betaine because of its strong antioxidant activity, leading to an anti-tumor effect. DOP can inhibit MNNG-induced PLGC models via regulating Wnt/β-catenin pathway and by changing endogenous metabolites.

Screening of Metabolic Markers of Qi‐Yin deficiency in Chronic Heart Failure Mice by Untargeted Metabolomics Method

Qi‐Yin deficiency is the main syndrome of chronic heart failure that one of the major heart disease that threatening human health. Clarifying the time‐history of Qi‐Yin deficiency in chronic heart failure disease, and further exploring the metabolic markers of Qi‐Yin deficiency can help to explain the pathological mechanism and Precise treatment of Qi‐Yin deficiency in chronic heart failure. This study adopted chronic heart failure model induced by coronary artery ligation in mice. We investigated the changes in the appearance of clinical indications corresponding to symptoms of Qi‐Yin deficiency at different time points, which aimed to clear the time‐history of Qi‐Yin deficiency appeared in chronic heart failure model mice. Positive (Yiqifumai) and negative (Danhong) control group were set up to assistant define the time‐history of Qi‐Yin deficiency. We found that Chronic heart failure mice appeared significant weight loss, decreased food intake, spontaneous activity , accelerated heart rate, and increased T wave amplitude in the first 2–3 weeks after modeling. What's more, Yiqifumai had better effects than Danhong in the improvement of Qi‐Yin deficiency in the first 2–3 weeks, which assistant proved that this period was Qi‐Yin deficiency. The coagulation function test results also suggest that it may turn into blood stasis syndrome around 4–5 weeks. In addition, LC‐MS/MS technology was used to determine the endogenous metabolites in serum and urine of mice on the positive ion mode. The data of accurate molecular weight and fragment ion peak were checked on HMDB database, and the identification methods of PCA and OPLS‐DA were used to analyze the multivariable data. Metabolites of the chronic heart failure model mice during the period of Qi‐Yin deficiency and non Qi‐Yin deficiency were compared to explore specific metabolite groups significant related to Qi‐Yin deficiency. The untargeted metabolomic workflow is showed in Figure 1. The results of metabolomic analysis of serum and urine of model mice showed that metabolites were well‐differentiated between the two groups at 2–3 weeks and 4–5 weeks. 29 significant difference metabolites in serum and 19 significant difference metabolites in urine were identified in this study. Hence, these results demonstrated that ischemia‐induced chronic heart failure may be mainly manifested as Qi‐Yin deficiency syndrome in the first 2–3 weeks, followed by conversion to blood stasis syndrome. Metabolites of chronic heart failure mice also changed as the syndrome transformed.Support or Funding InformationNational Natural Science Foundation of China (NO. 81774150)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

1H NMR-based serum metabolomics analysis of the age-related metabolic effects of perinatal exposure to BPA, BPS, BPF, and BPAF in female mice offspring.

The widespread application of bisphenols (BPs) in the industry has made them ubiquitous in the environment, causing potential environmental risks. Its unknown impacts on human being have received more and more attention. In this study, we have assessed the metabolic effects of perinatal exposure to bisphenol A (BPA) and its substitutes (bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF)) in female mice adolescent offspring and female mice adulthood offspring. 1H NMR-based serum metabolomics showed that metabolic profiles were disturbed with BPA and its three substitutes exposure in female mice adolescent offspring and female mice adulthood offspring. In addition, age-related metabolic effects were found based on changes in serum endogenous metabolites and metabolic pathways. Specifically, metabolic pathway analysis showed that major disturbed metabolic pathways in female mice adulthood offspring compare with female mice adolescent offspring also changed significantly. With the increase of age of the female mice offspring, changes in the metabolic pathways became more obvious in the BPA treatment group. Conversely, partially disturbed metabolic pathways were restored in the BPS, BPF, and BPAF treatment groups. In conclusion, perinatal exposure to BPA and its three substitutes significantly interferes with metabolic profiles and metabolic pathways, and this metabolic effects were age-related. These results offer more detailed information about the age-related metabolic effects of perinatal exposure to BPA, BPS, BPF, and BPAF in female mice offspring and provide data for systematic evaluation of the health risk assessment of BPA and its substitutes.

1H NMR toxicometabolomics following cisplatin-induced nephrotoxicity in male rats.

Cisplatin (CP) is an anti-cancer drug used for treatment of solid tumors, but the major adverse effect is drug-induced nephrotoxicity. The current study aimed to determine biomarkers that might predict nephrotoxicity induced by CP using serum or urinary proton nuclear magnetic resonance (1H NMR) spectral data in male Sprague-Dawley (S-D) rats. CP (0, 0.5 or 5 mg/kg) was intraperitoneally (i.p.) administered for single dose. Animals were sacrificed 2 days (D2) or 8 days (D8) after administration of CP in order to perform analysis of serum biochemistries and histopathologic examination. Urine samples were collected every 24 hr from pre-treatment to sacrifice. Serum and urinary 1H NMR spectral data revealed apparent differential clustering between control and CP-treated groups as evidenced by principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) in global and targeted profiling. The concentrations of endogenous serum metabolites, alanine, betaine, glucose, glutamine, lactate, and leucine were significantly increased on D2. Urinary concentrations of alanine, glucose, glycine, guanidinoacetate, acetate, and lactate were significantly elevated on D2 or D8, whereas concentrations of urinary metabolites, citrate and hippurate were significantly decreased on D2 or D8. The correlation of serum and urinary 1H NMR OPLS-DA with serum biochemistry and renal histopathologic changes suggests that 1H NMR urinalysis may be used to reliably predict or screen CP-induced nephrotoxicity. Data suggest that these altered endogenous metabolites might serve as specific biomarkers for CP-induced nephrotoxicity.