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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 166-174, 2024.
Artigo em Chinês | WPRIM | ID: wpr-1005266

RESUMO

ObjectiveMetabolomics was used to reveal the mechanism of Aconiti Lateralis Radix Praeparata(ALRP) in attenuating toxicity by processing from the aspects of amino acid metabolism, oxidative stress and energy metabolism by analyzing multiple metabolic pathways. MethodTwenty-four rats were randomly divided into control group, raw group and processed group, 8 rats in each group. The raw and processed group were given with 0.64 g·kg-1 of raw ALRP and processed ALRP respectively every day, the control group was given with an equal amount of normal saline once a day. After continuous administration for 7 days, the urine, serum and heart tissue of rats were collected. Pathological examination of the heart was carried out using hematoxylin-eosin(HE) staining, and the activities of lactate dehydrogenase(LDH) and creatine kinase-MB(CK-MB) in serum and cardiac tissues were detected by microplate assay and immunoinhibition assay. The effects of ALRP on rat heart before and after processing were compared and analyzed. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to perform urine metabolomics analysis, and multivariate statistical analysis was used to screen for differential metabolites related to ALRP in attenuating toxicity by processing, and pathway enrichment analysis was carried out to explore the processing mechanism. ResultHE staining showed that no obvious pathological changes were observed in the heart tissue of the control group, while obvious infiltration of inflammatory cells such as plasma cells and granulocytes was observed in the heart tissue of the raw group, indicating that the raw ALRP had strong cardiotoxicity. There was no significant difference in HE staining of heart tissue between the processed group and the control group, indicating that the toxicity of ALRP was significantly reduced after processing. Compared with the control group, the activities of LDH and CK-MB were significantly increased in serum and heart tissue of the raw group, and those were significantly decreased in serum and heart tissue of the processed group, suggesting that the myocardial toxicity of processed ALRP was reduced. A total of 108 endogenous differential metabolites associated with the raw ALRP were screened using multivariate statistical analysis in positive and negative modes, of which 51 differential metabolites were back-regulated by the processed ALRP. Biological analysis of the key regulatory pathways and associated network changes showed that the pathways related to toxicity of ALRP mainly included tryptophan metabolism, arginine and proline metabolism, phenylalanine metabolism, aminoacyl-tRNA biosynthesis, alanine, aspartate and glutamate metabolism, etc. The metabolic pathways related to the attenuation of processed ALRP mainly included aminoacyl-tRNA biosynthesis, tryptophan metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and caffeine metabolism. ConclusionThe processing technology of ALRP in Guilingji can significantly attenuate the cardiotoxicity of raw products, the mechanism mainly involves amino acid metabolism, oxidative stress and energy metabolism, which can provide experimental bases for the research related to the mechanism of toxicity reduction of ALRP by processing and its clinical safety applications.

2.
China Journal of Chinese Materia Medica ; (24): 5632-5640, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1008760

RESUMO

This study aimed to investigate the mechanism of Xihuang Pills in improving hyperplasia of mammary gland(HMG) in rats based on urine metabolomics using ultra-performance liquid chromatography-quadrupole-Orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS). The HMG rat model was established by intramuscular injection of estradiol benzoate solution(0.5 mg·kg~(-1), 25 days) followed by progesterone injection(5 mg·kg~(-1), 5 days). UPLC-Q-Orbitrap-MS technology was used to establish the endogenous small-molecule metabolic profiles in urine samples of rats in the blank group, the HMG model group, and Xihuang Pills group. Multivariate statistical analysis was performed for pattern recognition, t test and variable importance in the projection(VIP) were used to screen potential biomarkers. The significantly changed differential metabolites were identified using the online database Human Metabolome Database(HMDB). Metabolic pathway enrichment analysis was conducted using the MetaboAnalyst 5.0 database. The results showed that 90 differential metabolites with significant changes(P<0.05) were identified between the blank group and the HMG model group using the HMDB. Among them, 48 metabolites significantly reverted(P<0.05) after administration of Xihuang Pills, which may be related to the regulatory effect of Xihuang Pills. Thirteen metabolic pathways significantly associated with HMG were identified when the differential metabolites were imported into the MetaboAnalyst 5.0 database, and Xihuang Pills could modulate seven of these pathways. These metabolic pathways mainly involved histidine metabolism, arginine and proline metabolism, β-alanine metabolism, glycine, serine and threonine metabolism, tryptophan metabolism, pyrimidine metabolism, and amino sugar and nucleotide sugar metabolism. This study utilized UPLC-Q-Orbitrap-MS and urine metabolomics technology to analyze the mechanism of Xihuang Pills in improving HMG, laying the foundation for further in-depth research.


Assuntos
Humanos , Ratos , Animais , Cromatografia Líquida de Alta Pressão/métodos , Hiperplasia , Metabolômica/métodos , Metaboloma , Biomarcadores/urina
3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 130-137, 2020.
Artigo em Chinês | WPRIM | ID: wpr-873291

RESUMO

Objective::To explore the effect of Dioscoreae Nipponicae Rhizoma extract (DNRe) on rats with acute gouty arthritis (AGA) based on urine metabolomics and to search for the related potential biomarkers and metabolic pathways. Method::Rat model of AGA induced by monosodium urate (MSU) was selected, 40 rats were randomly divided into the blank group (k), the DNRe group (g), the model group (m), and the DNRe treatment group (gm), with 10 rats in each group. The drug-administered group was administered with DNRe at a dose of 0.48 g·kg-1 once a day for 5 days. The urine was gathered after the last administration, and analyzed with UPLC-Q-TOF/MS coupled with pattern recognition techniques, electrospray ionization (ESI) under positive and negative ion scanning mode was adopted, data collection range was m/z 100-1 500 with full scanning mode. Result::A total of 12 common potential biomarkers were identified as sarcosine, dimethylglycine, deoxycytidine, uric acid, 5-hydroxytryptamine (5-HT), L-cystathionine, 4-pyridoxic acid, deoxyuridine, melatonin, 5-methoxytryptamine, fumaric acid and cytidine. Compared with the blank group, the 12 potential biomarkers in the DNRe group were significantly down-regulated. Compare with the model group, 10 metabolites were up-regulated and 2 metabolites were down-regulated in the 12 potential biomarkers of the DNRe treatment group, the abnormal expression of 10 markers including sarcosine, uric acid, L-cystathionine, 4-pyridoxic acid, deoxyuridine, 5-methoxytryptamine, cytidine, dimethylglycine, melatonin, fumaric acid could be modulated by DNRe. The strongest metabolic pathways associated with AGA were cysteine and methionine metabolism, and tryptophan metabolism. Conclusion::The effect of DNRe on AGA may be related to the promotion of conversion level from cystathionine to cysteine in the cysteine and methionine metabolism, and the up-regulating melatonin level in tryptophan metabolism.

4.
Chinese Journal of Applied Physiology ; (6): 340-344 349, 2018.
Artigo em Chinês | WPRIM | ID: wpr-773748

RESUMO

OBJECTIVE@#To study the differential metabolites in urine and the characteristics of metabolic pathway of middle school students with chronic fatigue syndrome (CFS) before and after exercise, and then explain the metabolic mechanism of CFS.@*METHODS@#Eight male middle school students (age:17-19) with CFS were selectedas the CFS group according to CFS screening criteria of the U.S. centers.At the same time, 8 male health students of the same age from the same school were selected as the control group. They were administrated to do one-time exercise on the improved Harvard step (up and down steps 30 times/min for 3minutes). Their urinewascollected before and after exercise, and the differential metabolitesin urine were detected by liquid chromatography-mass spectrometry (LC-MS). The multidimensional statistical methods were used to analyze the metabolites by principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA). Finally, MetPA database was used to analyze the metabolites and to construct the correlativemetabolic pathways.@*RESULTS@#Compared with the control group, the creatine, indoleacetaldehyde, phytosphingosine and pyroglutamic acid were selected as differential metabolites and the contents of those were decreased significantly (<0.05 or <0.01) in CFS groupbefore the step movement. However, 11 differential metabolitesin CFS group were selected out after exercise, which were nonanedioic acid, methyladenosine, acetylcarnitine, capric acid, corticosterone, creatine, levonorgestrel, pantothenic acid, pyroglutamic acid, xanthosine and xanthurenic acid in sequence, the contents of methyladenosine and creatinewere significantly increased (<0.05) and the contents of the other 9 differentialmetabolites were significantly decreased (<0.05 or <0.01)compared with the control group.The 15 differential metabolites mentioned above were input MetPA database in order to analyze the metabolic pathways weighted score.The results showed that the arginine-proline metabolism pathway disordersweredetected in theCFS group before exercise, the marker metabolite wascreatine. And 3 metabolic pathwaysdisorder weredetectedin the CFS groupafter exercise, which were arginine-proline metabolism, biosynthesis of pantothenic acid and CoA, steroid hormone biosynthesis, and the marker metabolites, in turn, werecreatine, pantothenic acid and corticosterone.@*CONCLUSIONS@#The disorder of arginine-proline metabolic pathway is detected in CFS middle school students before exercise intervention. After exercise, it can be detected that the steroid hormone biosynthetic metabolic pathway, pantothenic acid and CoA metabolic pathways also have metabolic disorders.


Assuntos
Adolescente , Humanos , Masculino , Adulto Jovem , Biomarcadores , Exercício Físico , Síndrome de Fadiga Crônica , Metabolômica , Estudantes
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