Jun-sheng Tian

Antidepressant-like effects of the fractions of Xiaoyaosan on rat model of chronic unpredictable mild stress.

ETHNOPHARMACOLOGICAL RELEVANCE Xiaoyaosan (XYS), composed of Radix Bupleuri, Radix Angelicae Sinensis, Radix Paeoniae Alba, Rhizoma Atractylodis Macrocephalae, Poria, Herba Menthae, Rhizoma Zingiberis Recens and Radix Glycyrrhizae, is a valuable traditional Chinese medicine (TCM) which is used for the treatment of depression in China. In the formula, Radix Bupleuri usually serves as the principal drug, Radix Angelicae Sinensis and Radix Paeoniae Alba serve as the ministerial drugs, Rhizoma Atractylodis Macrocephalae, Poria, Herba Menthae and Rhizoma Zingiberis Recens serve as adjunctive drugs, Radix Glycyrrhizae serves as messenger drug, they coordinate with each other and enhance the effect of the formula. In our previous experiments, the antidepressant effect of XYS was revealed. However, the antidepressant part (or component) of this prescription was still obscure. We divided the XYS into five different polar fractions, and explored the antidepressant activity of five different polar fractions to identify the active fraction. MATERIALS AND METHODS Behavior research and metabonomics method based on (1)H NMR were used for efficacy study of different fractions in chronic unpredictable mild stress (CUMS) model of depression. Rats were divided into 8 groups and drugs were administered during the 21 days model building period. The urine samples of rats were collected overnight (12h) on 21 day and the metabolic profiling of the urine was measured using NMR. Multivariate analysis was also utilized to evaluate the active fraction of XYS. RESULTS In the behavior research, there were significant difference between the lipophilic fraction group (XY-A) and the model group. In addition, with pattern recognition analysis of urinary metabolites, the results showed a clear separation of the model group and control group, while XY-A group was much closer to the control group in the OSC-PLS score plot. Seven endogenous metabolites contributing to the separation of the model group and control group were detected, while XY-A group regulated the 5 perturbed metabolites showing a tendency of recovering to control group. CONCLUSIONS The present work suggested that petroleum ether fraction was the most effective fraction, implying that lipophilic components contribute to the antidepressant effect of XYS.

Dynamic analysis of the endogenous metabolites in depressed patients treated with TCM formula Xiaoyaosan using urinary 1H NMR-based metabolomics

ETHNOPHAMACOLOGICAL RELEVANCE Xiaoyaosan (XYS), one of the best-known traditional Chinese medicine prescriptions with a long history of use, is composed of Bupleurum chinense DC., Paeonia lactiflora Pall., Poria cocos (Schw.) Wolf, Angelica sinensis (Oliv.) Diels, Zingiber officinale Rosc., Atractylodes macrocephala Koidz., Glycyrrhiza uralensis Fisch., and Mentha haplocalyx Briq. For centuries, XYS has been widely used in China for the treatment of mental disorders such as depression. However, the complicated mechanism underlying the antidepressant activity of XYS is not yet well-understood. This understanding is complicated by the sophisticated pathophysiology of depression and by the complexity of XYS, which has multiple constituents acting on different metabolic pathways. The variations of endogenous metabolites in depressed patients after administration of XYS may help elucidate the anti-depressant effect and mechanism of action of XYS. The aim of this study is to establish the metabolic profile of depressive disorder and to investigate the changes of endogenous metabolites in the depressed patients before and after the treatment of Xiaoyaosan using the dynamic analysis of urine metabolomics profiles based on (1)H NMR. MATERIALS AND METHODS Twenty-one depressed patients were recruited from the Traditional Chinese Medicine Department of the First Affiliated Hospital of Shanxi Medical University. Small endogenous metabolites present in urine samples were measured by nuclear magnetic resonance (NMR) and analyzed by multivariate statistical methods. The patients then received XYS treatment for six weeks, after which their Hamilton Depression Scale (HAMD) scores were significantly decreased compared with their baseline scores (p≤0.01). Eight components in urine specimens were identified that enabled discrimination between the pre- and post-XYS-treated samples. RESULTS Urinary of creatinine, taurine, 2-oxoglutarate and xanthurenic acid increased significantly after XYS treatment (p≤0.05), while the urinary levels of citrate, lactate, alanine and dimethylamine decreased significantly (p≤0.05) compared with pre-treatment urine samples. These statistically significant perturbations are involved in energy metabolism, gut microbes, tryptophan metabolism and taurine metabolism. CONCLUSIONS The symptoms of depression had been improved after 6 weeks׳ treatment of XYS according to evaluation of HAMD scores. The dynamic tendency of the 8 metabolites that changed significantly during the treatment of XYS is consistent with the improvement in symptoms of depression. These metabolites may be used as biomarkers for the diagnosis of depressive disorders or the evaluation of the antidepressant as well as the exploration of the mechanism of depression.

1H-NMR-Based Metabonomic Studies on the Anti-Depressant Effect of Genipin in the Chronic Unpredictable Mild Stress Rat Model

The purpose of this work was to investigate the anti-depressant effect of genipin and its mechanisms using 1H-NMR spectroscopy and multivariate data analysis on a chronic unpredictable mild stress (CUMS) rat model. Rat serum and urine were analyzed by nuclear magnetic resonance (NMR)-based metabonomics after oral administration of either genipin or saline for 2 weeks. Significant differences in the metabolic profile of the CUMS-treated group and the control group were observed, which were consistent with the results of behavioral tests. Metabolic effects of CUMS included decreases in serum trimetlylamine oxide (TMAO) and β-hydroxybutyric acid (β-HB), and increases in lipid, lactate, alanine and N-acetyl-glycoproteins. In urine, decreases in creatinine and betaine were observed, while citrate, trimethylamine (TMA) and dimethylamine were increased. These changes suggest that depression may be associated with gut microbes, energy metabolism and glycometabolism. Genipin showed the best anti-depressive effects at a dose of 100 mg/kg in rats. These results indicate that metabonomic approaches could be powerful tools for the investigation of the biochemical changes in pathological conditions or drug treatment.

Analysis of the restorative effect of Bu-zhong-yi-qi-tang in the spleen-qi deficiency rat model using 1H-NMR-based metabonomics

ETHNOPHARMACOLOGICAL RELEVANCE Bu-zhong-yi-qi-tang (BT) is a classical formula for the treatment of spleen-qi descending, visceroptosis with hyposplenic qi, uterine prolapse, and rectal prolapse due to chronic diarrhea in traditional Chinese medicine (TCM) and has been identified as an effective drug for the treatment of TCM spleen-qi deficiency in clinical practice. The present study aimed to investigate the restorative effect and the potential mechanisms of Bu-zhong-yi-qi-tang in a rat spleen-qi deficiency model using (1)H-NMR-based metabonomics. MATERIALS AND METHODS The rat spleen-qi deficiency model was established as follows: oral administration of Radix Rhei extract (equivalent to 10g/kg body weight of the crude drug), loaded swimming, and starvation for 24h. Each of these treatments was administered consecutively every three days. Sixty male SD rats were randomly divided into five groups, and three of the groups received a different oral dose of the aqueous extract of Bu-zhong-yi-qi-tang during the last seven days of the three-week experimental period. The body weight and motor behavior of the rats were measured and recorded once a week. The endogenous metabolites in the plasma were analyzed using NMR in conjunction with multivariate and statistical techniques. In addition, the liver and spleen were removed and weighed. RESULTS All of the rats in the spleen-qi deficiency group presented pasty loose stools, inactiveness, grouping, a decrease in swimming endurance, and lackluster, loose, and disorderly behavior in addition to a significant decrease in body weight, spleen weight, and liver weight. In contrast, the abovementioned demonstrations were reversed to a certain extent in the rats treated with Bu-zhong-yi-qi-tang compared with the model group (p<0.05, p<0.01). A significant separation was determined between the control and model groups in the PCA score plot, which indicates that the spleen-qi deficiency model was successfully duplicated. The changes in the levels of endogenous metabolites in the plasma included lower levels of valine, leucine, and O-acetyl-glycoprotein and a higher concentration of lactate in the spleen-qi deficiency group compared with the control group. Treatment with Bu-zhong-yi-qi-tang at least partially returned the levels of these metabolites to the normal levels. CONCLUSIONS The restorative effects of Bu-zhong-yi-qi-tang in rats with spleen-qi deficiency were confirmed, and four endogenous metabolites were identified as potential biomarkers of the symptoms of spleen-qi deficiency and most likely play roles in the changes observed in certain metabolic pathways, such as the energy, protein, and glycolytic metabolisms.

A 1H-NMR plasma metabonomic study of acute and chronic stress models of depression in rats

To investigate and compare the metabonomic profiles of three stress-based models of depression, the effects of acute and chronic stress on the production of systemic endogenous metabolites were investigated. Such metabonomic analysis may provide researchers a new way of selecting appropriate animal models for the study of depression and antidepressants. Rats were subjected to one of three stress-based models: CUMS, FST-1d, or FST-14d. Endogenous metabolites excreted in plasma were analyzed using NMR in conjunction with multivariate and statistical techniques. The metabonomic study indicated that the concentration of different plasma metabolites could be used to differentiate among depression models: TMA, aspartic acid, glutamate, AcAc, NAc, alanine, lactate, Leu/Ile, lipids increased and proline, β-HB, valine decreased in the CUMS model; TMA decreased in the FST-1d model; α-glucose, β-glucose, β-HB, valine and lipids increased in the FST-14d model. The results suggested that metabonomics is a potentially appropriate method for evaluating depression models. According to the metabonomics study, CUMS model was more suitable and sensitive than the acute FST-1d model and predictable FST-14d model. The CUMS model was more appropriate for investigating both the efficacy of antidepressants and their mechanisms of action, while the FST-14d model should only be used for evaluating the efficacy of treatment.

Plasma-metabolite-biomarkers for the therapeutic response in depressed patients by the traditional Chinese medicine formula Xiaoyaosan: A 1H NMR-based metabolomics approach

BACKGROUND Depression is one of the most prevalent and serious mental disorders. Xiaoyaosan, a well-known Chinese prescription, has been widely used for the treatment of depression in China. Both clinical studies and animal experiments indicate that Xiaoyaosan has an obvious antidepressant activity. Additionally, a large number of candidate biomarkers have emerged that can be used for early disease detection and for monitoring ongoing treatment response to therapy because of their correlations with the characteristics of the disease. However, there have been few reports on biomarkers that measure the treatment response to the clinical use of Xiaoyaosan using a metabolomics approach. The current study is aimed at discovering biomarkers and biochemical pathways to facilitate the diagnosis of depression and the efficient evaluation of Xiaoyaosan using plasma metabolomics profiles based on (1)H NMR. METHODS Sixteen depressed patients diagnosed by standard methods (HAMD and CGI-SI) and sixteen healthy volunteers were recruited. (1)H NMR-based metabolomics techniques and multivariate statistical methods were used to analyze the plasma metabolites of the depressed patients before and after treatment and to compare them with healthy controls. RESULTS The plasma levels of trimethylamine oxide, glutamine and lactate in depressed patients increased significantly (p≤0.05) compared with healthy controls, whereas the levels of phenylalanine, valine, alanine, glycine, leucine, citrate, choline, lipids and glucose decreased significantly (p≤0.05). Additionally, alanine, choline, trimethylamine oxide, glutamine, lactate and glucose were returned to normal levels after Xiaoyaosan treatment. These statistically significant perturbations are involved in energy metabolism, amino acid metabolism and gut microbiota metabolism. LIMITATIONS Additional experimentation with gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS) is required to confirm our findings. CONCLUSIONS Application of these biomarkers in clinical practice may help to optimize the diagnosis of depression and to evaluate the efficacy of Xiaoyaosan. Metabolomics is promising as a biomarker discovery tool.

A GC–MS urinary quantitative metabolomics analysis in depressed patients treated with TCM formula of Xiaoyaosan

Xiaoyaosan, one of the best-known traditional Chinese medicine prescriptions, has been widely used in China for the treatment of mental disorders such as depression. Although both clinical application and animal experiments indicate that Xiaoyaosan has an obvious antidepressant effect, the mechanism still remains unclarified, and there are few studies quantitatively measured the biomarkers of Xiaoyaosan treatment by metabolomics to determination. In this study, 25 depressed patients and 33 healthy volunteers were recruited. A GC-MS based metabolomics approach and the multivariate statistical methods were used for analyzing the urine metabolites of depressed patients before and after treatment compared with healthy controls. Then the biomakers through metabolomics determination were carried out the quantitative analysis. In total, 5 metabolites were identified as the potential diseased and therapeutic biomarkers of depression and Xiaoyaosan. Alanine, citrate and hippurate levels were significantly increased in the urine samples from depressed patients compared with healthy controls, while phenylalanie and tyrosine levels were significantly decreased. However, after Xiaoyaosan treatment for 6 weeks, phenylalanie and tyrosine levels were significantly increased (p<0.05) and alanine, citrate and hippurate levels significantly decreased (p<0.05). Xiaoyaosan has a good priority on the treatment of depression and the ability to adjust the neurotransmitters to obtain the best treated response and also could regulate the metabolism of amino acids and promote to produce energy meet the needs of the body.

Combining biochemical with (1)H NMR-based metabolomics approach unravels the antidiabetic activity of genipin and its possible mechanism.

Diabetes mellitus is a typical heterogeneous metabolic disorder characterized by abnormal metabolism of carbohydrates, lipids and proteins. Genipin possesses a wide spectrum of biological activities including ameliorating effects on diabetes, but the definite mechanism of this effect remains unknown. To investigate the antidiabetic activities of genipin and explore the biochemical changes of serum endogenous metabolites on diabetic rats induced by alloxan, (1)H NMR spectroscopy coupled with multivariate data analysis was used to. All rats were randomly divided into six groups including negative control (NC) group, diabetic mellitus (DM) group, metformin hydrochloride group, high dose group of genipin, middle dose group of genipin and low dose group of genipin. Diabetes was induced by a single intraperitoneal injection of 120mg/kg body weight of alloxan. Serum samples were collected for the (1)H NMR-based metabolomics and clinical biochemical analysis. Daily oral administration of genipin (25, 50 and 100mg/kg body weight) and metformin hydrochloride (125mg/kg) for two weeks showed beneficial effects on blood glucose level (P<0.01). Significant differences in the metabolic profile as well as the result of biochemical parameters between the diabetic group and the control group were observed. The PLS-DA scores and corresponding loading plots demonstrated that genipin significantly restored the abnormal metabolic state. Detailed analysis of the altered metabolite levels indicated that genipin significantly ameliorated the disturbance in glucose metabolism, tricarboxylic acid cycle, lipid metabolism and amino acid metabolism. Genipin showed the best anti-diabetic effects at a dose of 100mg/kg in rats. This finding indicates that chemical and metabolomic approaches could be powerful tools for the investigation of the biochemical changes in pathological conditions or drug treatment.

Discovery, screening and evaluation of a plasma biomarker panel for subjects with psychological suboptimal health state using (1)H-NMR-based metabolomics profiles.

Individuals in the state of psychological suboptimal health keep increasing, only scales and questionnaires were used to diagnose in clinic under current conditions, and symptoms of high reliability and accuracy are destitute. Therefore, the noninvasive and precise laboratory diagnostic methods are needed. This study aimed to develop an objective method through screen potential biomarkers or a biomarker panel to facilitate the diagnosis in clinic using plasma metabolomics. Profiles were based on H-nuclear magnetic resonance (1H-NMR) metabolomics techniques combing with multivariate statistical analysis. Furthermore, methods of correlation analysis with Metaboanalyst 3.0 for selecting a biomarker panel, traditional Chinese medicine (TCM) drug intervention for validating the close relations between the biomarker panel and the state and the receiver operating characteristic curves (ROC curves) analysis for evaluation of clinical diagnosis ability were carried out. 9 endogenous metabolites containing trimethylamine oxide (TMAO), glutamine, N-acetyl-glycoproteins, citrate, tyrosine, phenylalanine, isoleucine, valine and glucose were identified and considered as potential biomarkers. Then a biomarker panel consisting of phenylalanine, glutamine, tyrosine, citrate, N-acetyl-glycoproteins and TMAO was selected, which exhibited the highest area under the curve (AUC = 0.971). This study provided critical insight into the pathological mechanism of psychological suboptimal health and would supply a novel and valuable diagnostic method.

1H NMR-based metabolic profiling of liver in chronic unpredictable mild stress rats with genipin treatment

Genipin, a hydrolyzed metabolite of geniposide extracted from the fruit of Gardenia jasminoides Ellis, has shown promise in alleviating depressive symptoms, however, the antidepressant mechanism of genipin remains unclear and incomprehensive. In this study, the metabolic profiles of aqueous and lipophilic extracts in liver of the chronic unpredictable mild stress (CUMS)-induced rat with genipin treatment were investigated using proton nuclear magnetic resonance ((1)H NMR) spectroscopy coupled with multivariate data analysis. Significant differences in the metabolic profiles of rats in the CUMS model group (MS) and the control group (NS) were observed with metabolic effects including decreasing in choline, glycerol and glycogen, increasing in lactate, alanine and succinate, and a disordered lipid metabolism, while the moderate dose (50mg/kg) of genipin could significantly regulate the concentrations of glycerol, lactate, alanine, succinate and the lipid to their normal levels. These biomakers were involved in metabolism pathways such as glycolysis/gluconeogensis, tricarboxylic acid (TCA) cycle and lipid metabolism, which may be helpful for understanding of antidepressant mechanism of genipin.