ABSTRACT
ObjectiveTo establish the identification method of Dalbergiae Odoriferae Lignum（DOL） and its counterfeits by nuclear magnetic resonance hydrogen spectrum（1H-NMR） combined with multivariate statistical analysis. Method1H-NMR spectra of DOL and its counterfeits were obtained by NMR， and the full composition information was established and transformed into a data matrix， and the detection conditions were as follows：taking dimethyl sulfoxide-d6（DMSO-d6） containing 0.03% tetramethylsilane（TMS） as the solvent， the constant temperature at 298 K（1 K=-272.15 ℃）， pulse interval of 1.00 s， spectrum width of 12 019.23 Hz， the scanning number of 16 times， and the sampling time of 1.08 s. Similarity examination and hierarchical cluster analysis（HCA） were performed on the data matrix of DOL and its counterfeits， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to analyze the data matrix and identify the differential components between them. In the established OPLS-DA category variable value model， the category variable value of DOL was set as 1， and the category variable value of the counterfeits was set as 0， and the threshold was set as ±0.3， in order to identify the commercially available DOL. The OPLS-DA score plot was used to determine the types of counterfeits in commercially available DOL， and it was verified by thin layer chromatography（TLC）. ResultThe results of similarity analysis and HCA showed that there was a significant difference between DOL and its counterfeits. OPLS-DA found that the differential component between DOL and its counterfeits was trans-nerolidol. The established category variable value model could successfully identify the authenticity of the commercially available DOL. The results of the OPLS-DA score plot showed that there were heartwood of Dalbergia pinnata and D. cochinchinensis in the commercially available DOL， and were consistent with the TLC verification results. ConclusionThere is a phenomenon that heartwood of D. pinnata and D. cochinchinensis are sold as DOL in the market. 1H-NMR combined with multivariate statistical analysis can effectively distinguish DOL and its counterfeits， which can provide a reference for the identification of them.
ABSTRACT
Objective:Metabolomics was used to analyze the dynamic changes of endogenous metabolites in serum and lung tissue of rats treated with water elution section of Siegesbeckiae Herba (SWES), and to explore the possible mechanism of lung injury and to search for the sensitive markers in serum and lung tissue. Method:SD rats were randomly divided into three groups, namely the normal group (normal saline), SWES high-dose group and SWES low-dose group (0.500, 0.125 g·kg-1·d-1). SWES was given for 4 weeks and stopped for 2 weeks. The weight and pathological examination were regarded as observation parameters. Serum and lung tissue samples were detected by nuclear magnetic resonance hydrogen spectrum (1H-NMR). The metabolites in rats were analyzed by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). Result:Lung inflammation was shown in SWES high-dose group and returned to normal after withdrawal for 2 weeks. The metabolic spectrum of SWES high-dose group was significantly different from the normal group. There were 11 metabolites were identified by 1H-NMR metabolomics, four differential metabolites were identified in serum [acetate, trimethylamine oxide (TMAO), glycine, myo-inositol] and nine differential metabolites were identified in lung tissue [lactate, acetate, phosphatidylcholine (PC), pyruvate, dimethylamine, glutamate, glycerophosphocholine (GPC), glycine, xanthine]. Lung injury was related to the disorder of pyruvate metabolism, glutamate metabolism and other pathways. Most of the metabolites in lung tissue had obviously levels of callback after drug withdrawal, which coincided with the pathological examination. Conclusion:The lung is one of the damaged organs caused by SWES, and the lung injury is reversible and may be related to energy metabolism and oxidative stress.
ABSTRACT
Objective:Metabolomics was used to analyze the brain tissue samples of model mice with chronic unpredictable mild stress (CUMS) depression, in order to find out the differential metabolites related to depression and to explore the possible antidepressant mechanism of iridoid part of Valerianae Jatamansi Rhizoma et Radix (IEFV). Method:Forty-two Kunming mice were randomly divided into 6 groups, including the normal group, the model group, the fluoxetine group (2.5 mg·kg-1) and the IEFV low, medium, and high dose groups (doses were 5.73, 11.47, 22.94 mg·kg-1, respectively). The behavioral and biochemical indicators of CUMS model mice were used for pharmacodynamic evaluation with IEFV and a positive drug (fluoxetine) as the intervention drugs. Then, the effect of IEFV on endogenous substances of the brain tissue in CUMS model mice were analyzed by nuclear magnetic resonance hydrogen spectrum (1H-NMR) metabolomics, and multivariate statistical analysis was used to identify the differential metabolites and to enrich the metabolic pathways involved in the differential metabolites. Result:After modeling, the immobility time of the model mice increased significantly, their sucrose preference rate and the excitatory neurotransmitters [5-hydroxytryptamine (5-HT) and norepinephrine (NE)] decreased significantly, indicating the success of modeling. The depression was relieved after IEFV administration, mainly manifested by the recovery of the immobility time, sucrose preference rate and the excitatory neurotransmitters (5-HT and NE). Principal component analysis (PCA) of endogenous metabolites in brain tissue showed that the model group could be significantly separated from the normal group, while the IEFV groups and fluoxetine group all showed a trend of deviating from the model group to the normal group, which was consistent with the behavioral results. The results of orthogonal partial least squares discriminant analysis (OPLS-DA) showed that there were 16 different metabolites between the model group and the normal group, including 12 water-soluble metabolites and 4 liposoluble metabolites. Seven potential metabolism pathways were obtained through MetPA analysis, including metabolism of phenylalanine, metabolism of phenylalanine, tyrosine and tryptophan, metabolism of taurine and hypotaurine acid, metabolism of alanine, aspartic acid and glutamic acid, biosynthesis of valine, leucine and isoleucine, metabolism of D-glutamine and D-glutamate and tricarboxylic acid cycle (TCA). IEFV-high dose group could significantly recall 11 differential metabolites. Conclusion:IEFV may play an antidepressant role mainly by affecting energy metabolism, amino acid metabolism and neurotransmitter levels, which provides a reference for further study on the antidepressant mechanism of IEFV.
ABSTRACT
Objective:The effect of processing on promoting digestion of Crataegi Fructus was investigated by the method of serum metabonomics based on 1H-NMR and the analysis of traditional biochemical indexes. Method:The dyspepsia model was induced by high protein and calorie diet. The small intestine propulsive rate in mice and the hormone level in rat gastrointestinal tract were used as pharmacodynamic indexes. After administration of Crataegi Fructus, fried and charred Crataegi Fructus, the changes of serum metabolites were analyzed by 1H-NMR technique combined with multivariate statistical analysis, so as to compare the therapeutic effects of each processed products on rats with dyspepsia model. Result:Each processed products group of Crataegi Fructus could improve the ability of intestinal propulsion and gastric emptying, especially in charred Crataegi Fructus group, caused by dyspepsia. The serum metabonomics proved that there were significant metabolic differences between the model group and the blank group. A total of 13 biomarkers related to dyspepsia, covering 3-hydroxybutyric acid, glycerophosphoryl choline (GPC), N-acetylglycoprotein, O-acetylglycoprotein, trimethylamine oxide (TMAO), alanine, acetic acid, glutamic acid, glutamine, creatine, leucine, lactic acid and glucose, were screened on the grounds of VIP value of S-plot, single factor ANOVA and area under receiver operating characteristic curve (ROC curve) (AUC). The metabolite composition of each administration group was the same except that the callback tendency of O-acetylglycoprotein, glutamine and GPC in charred Crataegi Fructus group was closer to that in the blank group. Conclusion:There are differences in the effect of promoting digestion to eliminate stagnation among processed products of Crataegi Fructus, charred Crataegi Fructus is more effective in improving dyspepsia, which may play a role in regulating gastrointestinal motility and energy metabolism.
ABSTRACT
Objective:Nuclear magnetic resonance hydrogen spectrum(1H-NMR) was adopted to investigate the effect of dry-cold environment and diet on urine and plasma metabolites of rats,in order to find possible biomarkers and their related metabolic pathways. Method:The dry-cold environment and diet were used as a stressors to intervene rats for 3 weeks,urine and plasma samples were collected.1H-NMR combined with multivariate data analysis were used to identify relevant metabolic markers and analyze their metabolic pathways. Result:The dry-cold environment and diet resulted in significant changes of 23 metabolites in urine and 15 metabolites in plasma;these metabolites were closely related to changes of various metabolic pathways,namely the tricarboxylic acid(TCA) cycle,pyruvate metabolism,glycolysis or gluconeogenesis,valine,leucine and isoleucine biosynthesis,glycine,serine and threonine metabolism,histidine metabolism;glyoxylic acid and dicarboxylic acid metabolism,alanine,aspartic acid and glutamate metabolism. Conclusion:Dry-cold environment and diet conditions can lead to the changes of various metabolic pathways in the body.The metabolomics based on 1H-NMR has the advantage of explaining the pathophysiological dynamics and overall changes of the body,which is of great significance for exploring the pathogenesis of non-deterministic disease factors.