Zhongfeng Li

Identification of metabolic biomarkers in patients with type 2 diabetic coronary heart diseases based on metabolomic approach

Type 2 diabetic coronary heart disease (T2DM-CHD) is a kind of serious and complex disease. Great attention has been paid to exploring its mechanism; however, the detailed understanding of T2DM-CHD is still limited. Plasma samples from 15 healthy controls, 13 coronary heart disease (CHD) patients, 15 type 2 diabetes mellitus (T2DM) patients and 28 T2DM-CHD patients were analyzed in this research. The potential biomarkers of CHD and T2DM were detected and screened out by 1H NMR-based plasma metabolic profiling and multivariate data analysis. About 11 and 12 representative metabolites of CHD and T2DM were identified respectively, mainly including alanine, arginine, proline, glutamine, creatinine and acetate. Then the diagnostic model was further constructed based on the previous metabolites of CHD and T2DM to detect T2DM-CHD with satisfying sensitivity of 92.9%, specificity of 93.3% and accuracy of 93.2%, validating the robustness of 1H NMR-based plasma metabolic profiling to diagnostic strategy. The results demonstrated that the NMR-based metabolomics approach processed good performance to identify diagnostic plasma biomarkers and most identified metabolites related to T2DM and CHD could be considered as predictors of T2DM-CHD as well as the therapeutic targets for prevention, which provided new insight into diagnosing and forecasting of complex diseases.

Metabolic effects of Hedyotis diffusa on rats bearing Walker 256 tumor revealed by NMR-based metabolomics.

Hedyotis diffusa, a traditional Chinese herbal medicine, is widely used for oncotherapy and shows a positive effect in the clinical treatment. But its mechanism of anticancer activities is complicated and unclear. This study was undertaken to assess the therapeutic effects and reveal detailed mechanisms of H. diffusa for oncotherapy. A Walker 256 tumor‐bearing rat model was established, and metabolomic profiles of plasma and urine were obtained from 1H NMR technique. Multivariate statistical analysis methods were used to characterize the discriminating metabolites between control (C), Walker 256 tumor‐bearing rats model (M), and H. diffusa treatment (H) groups. Finally, 13 and 10 metabolomic biomarkers in urine and plasma samples were further identified as characteristic metabolites in M group, whereas H group showed a partial metabolic balance recovered, such as ornithine, N‐acetyl‐l‐aspartate, l‐aspartate, and creatinine in urine samples, and acetate, lactate, choline, l‐glutamine, and 3‐hydroxybutyrate in plasma samples. On the basis of the methods above, we hypothesized H. diffusa treatment reduced the injury caused by Walker 256 tumor and maintained a metabolic balance. Our study demonstrated that this method provided new insights into metabolic alterations in tumor‐bearing biosystems and researching on the effects of H. diffusa on the endogenous metabolism in tumor‐bearing rats.

1H NMR-Based Metabolomics Study of the Toxicological Effects in Rats Induced by “Renqing Mangjue” Pill, a Traditional Tibetan Medicine

“RenqingMangjue” pill (RMP), as an effective prescription of Traditional Tibetan Medicine (TTM), has been widely used in treating digestive diseases and ulcerative colitis for over a thousand years. In certain classical Tibetan Medicine, heavy metal may add as an active ingredient, but it may cause contamination unintentionally in some cases. Therefore, the toxicity and adverse effects of TTM became to draw public attention. In this study, 48 male Wistar rats were orally administrated with different dosages of RMP once a day for 15 consecutive days, then half of the rats were euthanized on the 15th day and the remaining were euthanized on the 30th day. Plasma, kidney and liver samples were acquired to 1H NMR metabolomics analysis. Histopathology and ICP-MS were applied to support the metabolomics findings. The metabolic signature of plasma from RMP-administrated rats exhibited increasing levels of glucose, betaine, and creatine, together with decreasing levels of lipids, 3-hydroxybutate, pyruvate, citrate, valine, leucine, isoleucine, glutamate, and glutamine. The metabolomics analysis results of liver showed that after RMP administration, the concentrations of valine, leucine, proline, tyrosine, and tryptophan elevated, while glucose, sarcosine and 3-hydroxybutyrate decreased. The levels of metabolites in kidney, such as, leucine, valine, isoleucine and tyrosine, were increased, while taurine, glutamate, and glutamine decreased. The study provides several potential biomarkers for the toxicity mechanism research of RMP and shows that RMP may cause injury in kidney and liver and disturbance of several pathways, such as energy metabolism, oxidative stress, glucose and amino acids metabolism.

NMR-based Metabolomic Techniques Identify the Toxicity of Emodin in HepG2 Cells

Emodin is a natural anthraquinone derivative that is present in various herbal preparations. The pharmacological effects of emodin include anticancer, hepatoprotective, anti-inflammatory, antioxidant and even antimicrobial activities. However, emodin also has been reported to induce hepatotoxicity, nephrotoxicity, genotoxicity and reproductive toxicity. The mechanism of emodin’s adverse effects is complicated and currently not well understood. This study aimed to establish a cell metabonomic method to investigate the toxicity of emodin and explore its potential mechanism and relevant targets. In the present study, metabonomic profiles of cell extracts and cell culture media obtained using the 1H NMR technique were used to assess emodin toxicity in HepG2 cells. Multivariate statistical analyses such as partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to characterize the metabolites that differed between the control and emodin groups. The results indicated that emodin resulted in differences in 33 metabolites, including acetate, arginine, aspartate, creatine, isoleucine, leucine and histidine in the cell extract samples and 23 metabolites, including alanine, formate, glutamate, succinate and isoleucine, in the cell culture media samples. Approximately 8 pathways associated with these metabolites were disrupted in the emodin groups. These results demonstrated the potential for using cell metabonomics approaches to clarify the toxicological effects of emodin, the underlying mechanisms and potential biomarkers. Our findings may help with the development of novel strategies to discover targets for drug toxicity, elucidate the changes in regulatory signal networks and explore its potential mechanism of action.

Urinary metabonomic study of patients with acute coronary syndrome using UPLC-QTOF/MS

This urinary metabonomic study aimed to identify the potential metabolic biomarkers in acute coronary syndrome (ACS) patients. Ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) was used to analyze the urine samples from ACS patients and healthy controls. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were applied to characterizing the endogenous metabolites and potential biomarker, respectively. Among twenty biomarkers that functioned in nine metabolic pathways, nine biomarkers were found up-regulated significantly, including of isobutyryl‑l‑carnitine, 3‑methylglutarylcarnitine, cinnavalininate, l‑tryptophan, 3‑methyldioxyindole, palmitic acid, N4‑acetylaminobutanal, 3‑sulfino‑l‑alanine and S‑adenosyl‑l‑homocysteine. The other eleven biomarkers were showed down-regulated, including of l‑lactic acid, trigonelline, nicotinuric acid, l‑alanine, d‑alanyl‑d‑alanine, creatine, N4‑acetylaminobutanoate, glutathionyl spermidine, 5‑methoxytryptamine, kynurenic acid and xanthurenic acid. This study also implied that fatty acid metabolism, fatty acid β‑oxidation metabolism, amino acid metabolism and TCA cycle played important roles in ACS. Therefore, urinary metabolomics may improve the diagnosis efficacy of ACS and make it more accurate and comprehensive for ACS diagnosis.

The effects of jolkinolide B on HepG2 cells as revealed by 1H-NMR-based metabolic profiling

Abstract Jolkinolide B (JB), which is isolated from the dried root of Euphorbia fischeriana Steud., has been reported to possess various therapeutic effects, such as treatment of edema and abdominal distention and protection against acute lung injury, and it has also been reported to have anti‐inflammatory even antitumor properties. Thus, JB has always been considered a promising anticancer drug candidate. In the current work, a cellular metabolomics evaluation based on the nuclear magnetic resonance (NMR) approach was applied to investigate the mechanism of JB in HepG2 cells. In addition, biological assays such as the MTT assay, DAPI staining and the Annexin V‐FITC/PI assay were implemented to evaluate cell viability and apoptosis in JB‐treated cells. Subsequently, we used multivariate statistical analyses, such as principal component analysis (PCA) and orthogonal projection to latent structure with discriminant analysis (OPLS‐DA) to identify metabolic biomarkers. In total, 36 metabolites in the cell extract samples and 30 metabolites in the cell culture media samples were clearly identified to be altered after the treatment. Variations in the specific metabolites suggested that HepG2 cells that were exposed to JB displayed the disordered effects in multiple metabolic pathways, such as the tricarboxylic acid cycle, amino acid metabolism, GSH synthesis and pyruvate metabolism. NMR‐based cell metabolomics provided a holistic method for the identification of JBs antitumor mechanisms and the exploration of its potential applications in preclinical and clinical studies. Graphical abstract Figure. No Caption available.

PLASMA METABOLOMICS REVEALS NOVEL METABOLIC MARKERS OF CHRONIC HEART FAILURE

Objectives Chronic heart failure (CHF) is a progressive clinical syndrome characterised by inability of the heart to adequately pump blood to meet metabolic demands of the body. There is intense interest in the identification of novel biomarkers which could improve the diagnosis of chronic heart failure. The overarching goal of the work discussed here was to apply a high-throughput approach, using 1H-NMR spectroscopy to identify novel plasma biomarkers and metabolic signatures underlying chronic heart failure. Methods Plasma samples from 30 patients with systolic heart failure (EF<40% plus signs and symptoms of failure) and 15 controls were analysed by nuclear magnetic spectroscopy. Each spectroscopy divided into regions of 0.005 ppm width was integrated. After processing the data, orthogonal partial least square discriminant analysis (OPLS-DA) was performed using SIMCA-P+ software (v11.5, Umetrics, Sweden). Results The score plot of OPLS-DA showed good separation between case and control on the level of metabolites. Several metabolites of chronic heart failure patients were altered, including the increased levels of lactate, creatine, proline, leucine, isoleucine, low-density lipoprotein, very-low-density lipoprotein and the decreased levels of histine, glucose, glutamate, valine as well as high-density lipoprotein. Multiple biochemical changes indicated dyslipidemia, oxidative stress and alteration of energy metabolism in chronic heart failure patients. Some compound could also discriminate between different stages of diseases. These findings revealed potential biological mechanisms underlying chronic heart failure. Conclusions The NMR-based metabolomics approach demonstrates good performance to identify the plasma metabolomic markers and provides new insights into metabolic process related to chronic heart failure.