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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 31-42, 2024.
Artículo en Chino | WPRIM | ID: wpr-999158

RESUMEN

ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties, the chemical components of Sojae Semen Nigrum(SSN) and Sojae Semen Praeparatum(SSP) were systematically compared and analyzed, and the main fermentation products in different fermentation time were quantitatively analyzed, so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used for the structural identification of the chemical constituents of SSN and SSP, and with the aid of Progenesis QI v2.3 software, the negative ion mode was employed for principal component analysis(PCA) pattern recognition, and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis(OPLS-DA) for two-dimensional data to obtain S-plot, and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC, and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period, and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate(PITC) pre-column derivatization and high performance liquid chromatography(HPLC) gradient elution, and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value(TAV). ResultA total of 19 kinds of differential components were screened out, mainly soybean saponins and isoflavones, and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP, glycoside bond hydrolysis mainly occurred, and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones, the content of flavor substances such as amino acids increased gradually. In the post-fermentation process, protein degradation mainly occurred, after 8 d of post-fermentation, the content of isoflavones was basically stable, while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP, such as the TAV of glutamate is always ahead of other flavor substances, and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation, combined with the traditional evaluation index of "fragrant and strong", it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant, delicious and slightly sweet", and at the same time, the post-fermentation index compounds such as glutamic acid, alanine and valine should be added as the quality control indicators of SSP, so as to standardize the production process and improve the quality of SSP.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 169-178, 2024.
Artículo en Chino | WPRIM | ID: wpr-1006568

RESUMEN

ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang(LJZT), and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), and the resulting compounds were identified by using databases, such as MassBank, PubChem, ChemSpider, Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform(TCMSP), and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT, including liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature, database and MS information, a total of 79 compounds were identified from LJZT, including 31 flavonoids, 15 terpenoids, 14 nitrogen-containing compounds, 6 phenylpropanoids, 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges, and the precision, stability, reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5, 2.602 1, 0.082 6, 0.128 1, 1.778 6, 0.015 7, 0.006 7, 0.030 4, 0.003 2 mg·g-1, respectively. ConclusionThe established method can quickly, sensitively and accurately analyze the chemical constituents in LJZT, clarify that the material basis of LJZT is mainly flavonoids, terpenoids and nitrogen-containing compounds, and simultaneously determine the contents of the 9 components, which can lay a foundation for the research on quality control, mechanism and clinical application of LJZT.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 118-125, 2024.
Artículo en Chino | WPRIM | ID: wpr-1006276

RESUMEN

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS), to evaluate the establishment of a mouse model of liver Yin deficiency by thyroid tablet suspension combined with 10% carbon tetrachloride(CCl4) from the perspective of non-targeted metabolomics, in order to lay the foundation for the establishment of a traditional Chinese medicine(TCM) syndrome model. MethodA total of 24 mice were randomly divided into blank group and model group. The model group was given thyroid tablet suspension(0.003 2 g·kg-1) by gavage for 14 consecutive days, and 10% CCl4(5 mL·kg-1) was intraperitoneally injected once a week to establish a liver Yin deficiency model, while the blank group was injected with an equal amount of olive oil intraperitoneally and gavaged with an equal amount of distilled water, and was fed with normal feed. After the modeling was completed, 6 mice in each group were randomly selected, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), cyclic adenosine monophosphate(cAMP), cyclic guanosine monophosphate(cGMP), interleukin(IL)-6, IL-10, tumor necrosis factor-α(TNF-α)were measured in the mice serum, and malondialdehyde(MDA), superoxide dismutase(SOD), total protein(TP), hydroxyproline(HYP) and other indicators were measured in the mice liver. Liver tissue sections were taken for hematoxylin-eosin(HE) staining and observing pathological changes. The remaining 6 mice in each group were subjected to UPLC-Q-TOF-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen differential metabolites in the liver Yin deficiency mouse model, Kyoto Encyclopedia of Genes and Genomes(KEGG) database was used to analyze the corresponding metabolic pathways of differential metabolites. ResultCompared with the blank group, mice in the model group showed liver Yin deficiency manifestations such as reduced body weight, fatigue and sleepiness, disheveled and lusterless hair, irritability. The levels of ALT, cAMP/cGMP, IL-6, AST, MDA, cAMP, TNF-α significantly increased(P<0.05, P<0.01), while the levels of SOD, IL-10 and cGMP significantly decreased(P<0.05, P<0.01), and the changes of HYP and TP were not statistically significant. Hepatic steatosis and distortion of the radial arrangement of the liver plate cells were seen in the section images of the model group, endogenous substances were clearly separated, and 252 differential metabolites were identified in the serum samples, which were mainly involved in the metabolic pathways of purine metabolism, steroid hormone biosynthesis and pyrimidine metabolism. A total of 229 differential metabolites were identified in the liver samples, mainly involving nucleotide metabolism, purine metabolism, steroid hormone biosynthesis, pyrimidine metabolism, antifolate resistance, insulin resistance, primary bile acid biosynthesis, prostate cancer, sulfur relay system, arachidonic acid metabolism and other metabolic pathways. ConclusionThe successful establishment of liver Yin deficiency model in mice by CCl4 combined with thyroid hormone is evaluated through the investigation of serum and liver metabolomics, combined with biochemical indicators, which provides a biological basis and experimental foundation for the Yin deficiency syndrome model of TCM.

4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 166-174, 2024.
Artículo en Chino | WPRIM | ID: wpr-1005266

RESUMEN

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.

5.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 124-132, 2024.
Artículo en Chino | WPRIM | ID: wpr-1003774

RESUMEN

ObjectiveTo analyze the antidepressant quality markers(Q-Marker) of Bupleuri Radix(BP) before and after vinegar-processing by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), multivariate statistical analysis and network pharmacology. MethodUPLC-Q-TOF-MS was used to analyze the chemical basis of raw and vinegar-processed products of BP, and principal component analysis(PCA) orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to identify the differential components in BP that changed significantly before and after vinegar-processing, which were regarded as candidate quality markers(Q-Marker). Then the disease-drug-component-target network related to antidepressant effect of BP was constructed by network pharmacology, and the antidepressant Q-Marker of raw and vinegar-processed products of BP was determined. Rats were randomly divided into blank group, model group, fluoxetine group(2.67 mg·kg-1) and total saponin group(0.72 mg·kg-1), except the blank group, rats in the other groups were subjected to chronic unpredictable mild stress(CUMS). Three weeks after the start of modeling, rats in each administration group were given the corresponding dose of drugs once a day for 4 weeks, and rats in the blank and model groups were given normal saline with dose of 10 mL·kg-1. At 1 day before modeling, 21 days and 28 days after administration, body mass weighing, sucrose preference test and open field test were performed on each group . After 28 days of administration, real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was used to detect the mRNA expression levels of phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), mammalian target of rapamycin(mTOR), glycogen synthase kinase-3β(GSK-3β), forkhead box transcription factor O3a(FoxO3a) and β-catenin in hippocampal tissues of rats in each group, while protein expression levels of PI3K, Akt, mTOR and FoxO3a in hippocampal tissues of rats in each group were detected by Western blot. ResultThere were 19 components in BP showed significant changes before and after vinegar-processing, and 9 components such as saikosaponin A, saikosaponin B1, saikosaponin B2, saikosaponin C and saikosaponin D were identified as potential Q-Marker through S-plot differential marker screening. Combined with the disease-drug-component-target network, saikosaponin A, saikosaponin B1, saikosaponin B2 and saikosaponin D were identified as antidepressant Q-Marker of raw and vinegar-processed products of BP. According to the results of pharmacodynamic tests, after 28 d of administration, compared with the blank group, the body mass, sucrose preference index and open field total score of rats in model group, fluoxetine group and total saponin group decreased significantly(P<0.01). Compared with the model group, the body mass, sucrose preference index and open field total score in total saponin group increased significantly(P<0.01). Compared with the blank group, mRNA expression levels of PI3K, Akt, mTOR and β-catenin in hippocampus of rats in the model group decreased significantly(P<0.05), while mRNA expression levels of GSK-3β and FoxO3a increased significantly(P<0.05). Compared with the model group, mRNA expression levels of PI3K, Akt, mTOR and β-catenin in hippocampus of rats in the total saponin group were increased significantly(P<0.05), while mRNA expression levels of GSK-3β and FoxO3a decreased significantly(P<0.05). Compared with the blank group, the protein expression levels of Akt and mTOR in hippocampus of the model group decreased significantly(P<0.01), while the protein expression levels of PI3K and FoxO3a increased significantly(P<0.01). Compared with the model group, the expression level of Akt in hippocampus of the total saponin group increased significantly(P<0.01), the mTOR expression level was increased but not statistically significant, while the protein expression levels of PI3K and FoxO3a decreased significantly(P<0.01). ConclusionThe chemical constituents of BP changed greatly after vinegar-processing, and the antidepressant Q-Marker of raw and vinegar-processed products of BP was determined by chemical basis, pharmacodynamics, network pharmacology and signaling pathway, which provided a reference for further research on quality control, pharmacodynamic substance basis and processing mechanism of BP.

6.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 179-188, 2024.
Artículo en Chino | WPRIM | ID: wpr-1011457

RESUMEN

ObjectiveTo clarify the differences in the efficacy and mechanism of different processed products of Atractylodes chinensis rhizoma by the pharmacodynamics and metabolomics studies of raw, bran-fried and rice water-processed products on rats with spleen deficiency. MethodSixty male SD rats were randomly divided into blank group, model group, raw product group(3.75 g·kg-1), bran-fried product group(3.75 g·kg-1), rice water-processed product group(3.75 g·kg-1) and Shenling Baizhusan group(6.7 g·kg-1), with 10 rats in each group. The method of excessive fatigue+improper diet was used to establish a spleen deficiency model in rats. After the end of modeling, except for the blank and model groups, each dosing group was given the corresponding drug suspension, the immune organ coefficients of each group of rats were examined, the levels of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), immunoglobulin G(IgG), amylase(AMS), motilin(MTL), gastrin(GAS), Na+-K+-adenosine triphosphatase(ATPase), aquaporin 2(AQP2), AQP3 and AQP8 in rats were measured by enzyme-linked immunosorbent assay(ELISA). Ultra high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS) combined with orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to search for biomarkers in the plasma samples of spleen-deficient rats by using two criteria[P<0.05 and variable importance in the projection(VIP) value>1], and to compare the different modulatory effects of the three decoction pieces on the splenic-deficient biomarkers, and metabolic pathway analysis was conducted through the Kyoto Encyclopedia of Genes and Genomes(KEGG) database. ResultCompared with the blank group, the thymus index and spleen index of rats in the model group were significantly decreased(P<0.05), the levels of IL-6, TNF-α, IgG and AQP2 were significantly increased(P<0.05), the levels of AMS, GAS, MTL, AQP3, AQP8 and Na+-K+-ATPase were significantly decreased(P<0.05). Compared with the model group, raw products, bran-fried products and rice water-processed products all increased thymus index and spleen index(P<0.05), decreased IL-6, TNF-α, IgG and AQP2 levels(P<0.05), and increased AMS, GAS, MTL, AQP3, AQP8 and Na+-K+-ATPase levels to different degrees. A total of 176 differential metabolites were screened in the model group compared with the blank group, of which 75, 72 and 84 biomarkers were called back by the raw products, bran-fried products and rice water-processed products, respectively(P<0.05, P<0.01). Raw products of A. chinensis rhizoma mainly affected glycine, serine and threonine metabolism. Bran-fried products mainly affected alanine, aspartate and glutamate metabolism, D-arginine and D-ornithine metabolism. Rice water-processed products mainly affected glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, citrate cycle, thiamine metabolism, D-arginine and D-ornithine metabolism. ConclusionRaw products, bran-fried products and rice water-processed products of A. chinensis rhizoma all have good spleen strengthening effects, among which the effects of bran-fried products and rice water-processed products were stronger. Meanwhile, raw products has the strongest dryness, followed by bran-fried products, and the weakest dryness of rice water-processed products. The three decoction pieces are able to significantly modulate metabolic abnormalities in spleen-deficient rats, and the mechanism may be related to amino acid metabolism such as glycine, serine and threonine metabolism as well as alanine, aspartate and glutamate metabolism.

7.
Acta Pharmaceutica Sinica ; (12): 2468-2475, 2023.
Artículo en Chino | WPRIM | ID: wpr-999138

RESUMEN

Dengzhan Shengmai capsule, as a compound Chinese patent medicine, consists of four herbs: Herba Erigerontis, Ginseng, Ophiopogon, and Schisandrae Chinensis Fructus, and contains significant components of flavonoids, lignans, saponins, and organic acids. It is widely used clinically to treat cerebrovascular diseases such as chronic cerebral hypoperfusion and dementia with remarkable efficacy. This study proposes a research strategy for multi-component traditional Chinese medicine metabolites based on prediction databases and unfolds the analysis using Dengzhan Shengmai capsule as an example. Using the UPLC-Q-TOF/MS method, the analytical method was established and detected biological samples such as urine, feces, and bile of rats before and after administration based on the prediction of theoretical metabolites of Dengzhan Shengmai capsule. The possible secondary fragment ion information of metabolites was identified by comparing the detected results with prediction databases. The metabolites were identified based on the archetypal component mass spectrometric cleavage law and multistage mass spectrometric data. 51 metabolites, mainly flavonoid, organic acid, and lignan constituents, were finally identified from rat biosamples based on 306 theoretical metabolites of Dengzhan Shengmai capsule. This study provides a new strategy for the identification of metabolites in vivo and the analysis of metabolic pathways of TCM. The study complied with the procedures established by the Animal Experiment Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and passed the animal experiment ethics examine (No. 00003645).

8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 140-149, 2023.
Artículo en Chino | WPRIM | ID: wpr-997667

RESUMEN

ObjectiveTo explore the material basis of bile-processed Coptidis Rhizoma clearing excessive fire of liver-gallbladder based on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF/MS) metabolomics and molecular docking. MethodUPLC-Q-TOF/MS metabolomics was used to analyze the chemical constituents of Coptidis Rhizoma, water-processed Coptidis Rhizoma and bile-processed Coptidis Rhizoma. Chromatographic separation was achieved with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase in gradient elution(0-2 min, 5%B; 2-20 min, 5%-65%B; 20-40 min, 65%-10%B; 40-45 min, 10%B; 45-46 min, 10%-95%B; 46-49 min, 95%B), and electrospray ionization(ESI) was applied and operated in positive and negative ion modes, the acquisition range was m/z 80-1 200. Based on this, partial least squares-discriminant analysis(PLS-DA) and variance analysis were used to screen the differential compounds among the three products of Coptidis Rhizoma. Network pharmacology and molecular docking were used to verify the degree of association between differential compounds and excessive fire of liver-gallbladder syndrome. ResultA total of 33 chemical constituents were identified, including 2 phenolic acids, 5 binding bile acids and 26 alkaloids. And 16 differential compounds were identified by multivariate statistical analysis, including 11 alkaloids and 5 binding bile acids. Pathway enrichment analysis in the Kyoto Encyclopedia of Genes and Genomes(KEGG) database yielded 8 pathways related to excessive fire of liver-gallbladder, and the key protein phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform(PIK3CA) was obtained according to the "component-target-pathway" network analysis. Molecular docking results showed that 11 alkaloids had good binding ability with PIK3CA. ConclusionPorcine bile is unique in the processing of bile-processed Coptidis Rhizoma, which can promote the production and dissolution of 11 alkaloids, including berberine and dihydrochelerythrine. Based on the results of molecular docking and reported pharmacological experiments, it can be concluded that 16 different compounds such as berberine, dihydrochelerythrine and taurohyodeoxycholic acid are the material basis of bile-processed Coptidis Rhizoma.

9.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 122-130, 2023.
Artículo en Chino | WPRIM | ID: wpr-997665

RESUMEN

ObjectiveBased on serum pharmacochemistry and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) the transitional components in the serum of rats after intragastric administration of water extract of Alismatis Rhizoma(AR)and salt-processed Alismatis Rhizoma(SAR) were compared. MethodSD rats were randomly divided into blank group, AR group(10 g·kg-1) and SAR group(10 g·kg-1), 3 rats in each group, the administration groups were given AR and SAR aqueous extracts by gavage, respectively, and the blank group was given an equal volume of drinking water by gavage once in the morning and once in the evening, for 3 consecutive days. Sixty minutes after the last administration, blood was collected from the eye orbits, and the serum samples were prepared. The serum samples were prepared on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm, 1.7 μm) with the mobile phase of acetonitrile(A)-0.1% formic acid aqueous solution(B) in a gradient elution(0-10 min, 10%-50% A; 10-27 min, 50%-95%A; 27-27.1 min, 95%-10% A; 27.1-30 min, 10%A), the data were collected at a flow rate of 0.3 mL·min-1 in positive ion mode with a scanning range of m/z 100-1 200. Based on the self-constructed chemical composition library of AR, the total ion flow diagrams and secondary MS fragmentation information of the aqueous extracts of AR and SAR, as well as the administered serum and the blank serum, were compared with each other by UNIFI 1.9.2, so as to deduce the possible blood-migrating constituents and their cleavage patterns in the aqueous extracts, and the response intensity ratios of each chemical component were calculated before and after processing. ResultA total of 20 components, including 5 prototypical components and 15 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of AR. And 14 components, including 5 prototypical components and 9 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of SAR. Of these, 13 components were common to both of them, including 5 prototypical components and 8 metabolites. The 5 prototypical components were 16-oxoalisol A, alisol A 24-acetate, alisol A, alisol B and alisol C. The metabolites were mainly involved in phase Ⅰ metabolism(oxidation) and phase Ⅱ metabolism(glucuronidation). There was a big change in the intensity of response of the common components before and after salt-processing, and the response intensities of the prototypical components, 16-oxoalisol A, alisol B and alisol C, were elevated, while the type and response intensity of metabolites were generally decreased, and it was hypothesized that the metabolic rate of terpenoids might be slowed down after salt-processing of AR, so that the blood-migrating constituents could participate in the metabolism of the body more in the form of prototypes. ConclusionSalt-processing of AR may promote the absorption of prototypical components into the blood by slowing down the metabolic rate of terpenoids, which can provide support for the research on material basis of AR and SAR.

10.
Acta Pharmaceutica Sinica ; (12): 1307-1316, 2023.
Artículo en Chino | WPRIM | ID: wpr-978691

RESUMEN

The goal of this work was to investigate the antidepressant fraction from Radix Paeoniae Alba and identify its major chemical constituents. Corticosterone injured rat phaeochromocytoma (PC12) cells and behavioral despair depression models of mice were used to evaluate the antidepressant effects of Radix Paeoniae Alba (Bai-Shao) ethanol extract (BS-E) and its three fractions (BS-10E, BS-60E, BS-95E) isolated by macroporous resin column chromatography. Animal experimental procedures were approved by the Animal Ethics Committee of the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College (approval No.: SLXD-20210618051). The results showed that BS-E, BS-10E and BS-60E had protective effects against PC12 cells injury induced by corticosterone, among which BS-60E had the strongest protective effect. BS-60E could significantly shorten the time of forced swimming and tail suspension in despair depression models of mice, and was identified as the antidepressant fraction of Radix Paeoniae Alba. The major chemical constituents in the antidepressant fraction were identified by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and their proposed fragmentation pathways in MS spectra were deduced. A total of 79 chemical constituents were identified from BS-60E, including 36 monoterpenes, 34 polyphenols, 6 oligosaccharides, and 3 other constituents, and monoterpenes and polyphenols may be major effective constituents of BS-60E.

11.
Acta Pharmaceutica Sinica ; (12): 1880-1893, 2023.
Artículo en Chino | WPRIM | ID: wpr-978661

RESUMEN

In this study, untargeted metabolomics technology based on ultra-high-performance liquid chromatography-quadrupole/time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze and identify the overall chemical components of Juniperri Caulis et Folium. Chemical markers for the identification of different Juniperri Caulis et Folium species were screened by integrated principal component analysis and partial least squares discriminant analysis. A total of 58 chemical components were detected and 46 of them were identified, including 26 flavonoids, 8 organic acids and their derivatives, 4 phenylpropanoids, 3 terpenoids, and 5 other components. Among them, methylsyringin and ekersenin were identified for the first time. In the positive ion mode, 12 markers were screened, and in the negative ion mode, 13 markers were screened for species identification. In summary, UPLC-Q-TOF-MS/MS metabonomics technology combined with chemometrics method can effectively reveal the chemical composition differences of different Juniperri Caulis et Folium species, and provide reference for its species identification and quality control.

12.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 166-173, 2023.
Artículo en Chino | WPRIM | ID: wpr-969612

RESUMEN

ObjectiveTaking the rat model of spleen-stomach damp-heat syndrome(SSDHS) as the research object, this study aimed to investigate the potential biomarkers of SSDHS and the related metabolic pathways based on urine metabolomics, and tried to reveal the essence of SSDHS at the level of endogenous small molecular metabolites. MethodSixteen SD rats were randomly divided into normal and model groups. The normal group was fed normal chow and the model group was fed with 200 g·L-1 honey water daily, and lard and Chinese Baijiu alternately on alternate days for 17 days. The SSDHS model rats were exposed to external dampness-heat environment with temperature at 30-34 ℃, relative humidity of 95% for 2 h at the same time every day from the 10th day for 7 d. Then, the model was evaluated by observing the general conditions of the rats, measuring the contents of motilin(MTL) and gastrin(GT) in plasma by enzyme-linked immunosorbent assay(ELISA), and examining the histopathology of gastronitestinal tissues. In additon, the urine metabolomics analysis was performed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), and the detection conditions was as follows:ACQUITY™ UPLC BEH C18 column(2.1 mm×100 mm, 1.7 μm), mobile phase of 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B) for gradient elution (0-3 min, 1%-18%B; 3-8 min, 18%-40%B; 8-10 min, 40%-100%B), the flow rate of 0.4 mL·min-1, electrospray ionization(ESI) in positive and negative ion modes, scanning range of m/z 50-1 000. The univariate and multivariate statistical analysis were constructed for screening inter-group differential ions, the element composition was calculated according to the precise relative molecular weight, and ion information was matched with databases such as Human Metabolome Database(HMDB) to identify biomarkers. Kyoto Encyclopedia of Genes and Genomes(KEGG) database was used to obtain the biological information of metabolites, and their associated metabolic pathways were analyzed by MetaboAnalyst 5.0. ResultCompared with the normal group, the rectal temperature of the model group increased significantly(P<0.01), the levels of plasma MTL and GT decreased significantly(P<0.05, P<0.01), and pathological changes such as bleeding, congestion and inflammatory infiltration in the gastric and colonic tissues. A total of 25 differential metabolites such as L-histidine, citric acid and isocitric acid were found to be the potential biomarker of SSDHS by urine metabolomics, 13 of which were phase Ⅱ metabolites of endogenous substances(glucuronic acid conjugates, sulfuric acid conjugates and acetyl conjugates), involving the metabolic pathways of histidine metabolism, tricarboxylic acid cycle, glyoxylate and dicarboxylate metabolism. ConclusionSSDHS primarily causes disorders of histidine metabolism, tricarboxylic acid cycle, glyoxylate and dicarboxylate metabolism, as well as the imbalance of the activation/inactivation of endogenous metabolites, which may involve the immune response, material and energy metabolism, inflammatory response and intestinal flora, and may provide a basis for the establishment and application of SSDHS model.

13.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 232-243, 2023.
Artículo en Chino | WPRIM | ID: wpr-965838

RESUMEN

ObjectiveUsing multi-omics technology, we conducted the present study to determine whether dexamethasone has therapeutic effect on pneumonia rats through the regulation of intestinal flora and metabolites. MethodsTotally 18 Sprague-Dawley rats were randomly divided into 3 groups (n = 6 each): Control group, Model group and Dexamethasone (Dex) group. Lipopolysaccharide (LPS) was continuously injected intraperitoneally into rats at a dose of 4 mg/kg for 7 days to induce pneumonia except the Control group. Then the Dex group was given Dex at a dose of 2 mg/kg via oral gavage for 12 days, and both the other two groups received continuously equal volume of sterile PBS buffer for 12 days. On the 19th day, lung, plasma, feces and intestinal contents of rat were collected. Hematoxylin-eosin (H&E) staining and Bio-plex suspension chip system were applied to evaluate the effect of Dex on pneumonia. Furthermore, metagenomic sequencing and UPLC-Q-TOF-MS/MS technology were employed to determine the intestinal flora and metabolites of rats, respectively. ResultsH&E staining results showed that the lung tissue of the Model group was infiltrated with inflammatory cells, the alveolar septum was increased, alveolar hemorrhage, and histological lesions were less severe in Dex group than in the model group. The levels of 3 inflammatory cytokines including TNF-α (P < 0.000 1), IL-1α (P = 0.009 6) and IL-6 (P < 0.000 1) in the Model group were increased compared with the Control group, while Dex treatment reduced the levels of the three inflammatory factors. Taken together, Dex treatment effectively reversed the features of pneumonia in rats. Metagenomic analysis revealed that the intestinal flora structure of the three groups of rats was changed. In contrast with the Model group, an increasing level of the Firmicutes and an elevated proportion of Firmicutes/Bacteroidetes were observed after Dex treatment. Dex-treated rats possessed notably enrichment of Bifidobacterium, Lachnospiraceae and Lactobacillus. Multivariate statistical analysis showed a great separation between Model group and Dex group, indicating metabolic profile changes. In addition, 69 metabolites (P < 0.05) were screened, including 38 up-regulated in the Model group and 31 elevated in the Dex group, all of which were mainly involved in 3 metabolic pathways: linoleic acid metabolism, tryptophan metabolism and primary bile acid biosynthesis. ConclusionsIn summary, we demonstrate the beneficial effects of Dex on the symptoms of pneumonia. Meanwhile, integrated microbiome-metabolome analysis reveals that Dex improves LPS-induced pneumonia in rats through regulating intestinal flora and host metabolites. This study may provide new insights into the mechanism of Dex treatment of pneumonia in rats.

14.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 202-209, 2023.
Artículo en Chino | WPRIM | ID: wpr-965664

RESUMEN

ObjectiveTo develop a quality control method for the simultaneous determination of multiple active components in Nymphaeae Flos aiming at the problems of the single index for quality control and the relatively low overall quality control level. MethodUltra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)was used to identify and select the index components for quality control with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B)for gradient elution (0-2 min, 3%-8%B; 2-4 min, 8%-10%B; 4-13 min, 10%-15%B; 13-19 min, 15%-20%B; 19-26 min, 20%-45%B) at a flow rate of 0.4 mL·min-1, detection wavelength of 350 nm, electrospray ionization(ESI), negative ion scanning mode, ion source temperature of 120 ℃, scanning range of m/z 100-1 200, transmit collision energy of 6 eV for low-energy scanning and 25-50 eV for high-energy scanning. High performance liquid chromatography(HPLC)was used to establish the quality control method for the simultaneous determination of multi-index components with the mobile phase of 0.2% phosphoric acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-30 min, 12%-15%B; 30-60 min, 15%-22%B; 60-90 min, 22%-40%B)and detection wavelength of 350 nm. The preparation method of the test solution for content determination was refluxing extraction for 60 min with 80 times the amount of 70% methanol. ResultBy comparing the retention time, ultraviolet absorption characteristics, MS and MS/MS spectrometric signals in the samples with the reference substances, 8 active components with high contents, including brevifolincarboxylic acid, ellagic acid, rutin, nicotiflorin, astragalin, quercetin, quercetin-3-methylether and kaempferol, were identified qualitatively from Nymphaeae Flos, which were selected as the index components for quality control. Under the established HPLC conditions, the above 8 components could be well separated(resolution>1.5), and showed good linearity(r=0.999 9)between the concentration ranges of 1.99-99.6, 1.76-176, 1.52-75.8, 3.60-180, 0.964-96.4, 1.18-118, 1.94-96.8, 1.04-104 mg·L-1 and the peak areas, respectively. The detection limits of them were 10-49 μg·L-1, and the limits of quantitation were 34-164 μg·L-1. The average recoveries were 97.12%-103.1% with the relative standard deviations (RSDs) were 1.1%-2.2%. ConclusionA quality control method for simultaneous determination of the multiple active components in Nymphaeae Flos have been developed, which is simple, accurate and reproducible, and it can provide a scientific basis for the formulation of quality standard of this herb and lay a research foundation for the transformation of Uygur hospital preparations containing Nymphaeae Flos into new drugs.

15.
Acta Pharmaceutica Sinica ; (12): 740-749, 2023.
Artículo en Chino | WPRIM | ID: wpr-965630

RESUMEN

The aim is to study the tissue distribution characteristics of eight effective components in normal rats after oral administration of Ziziphi Spinosae Semen (ZSS) aqueous extract. An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) analysis method was developed and validated for the determination of four flavonoids and four saponins in rat tissue using puerarin and ginsenoside Re as the internal standard (IS), respectively. Tissue samples including the heart, liver, spleen, lung, kidney, muscle, brain, small intestine, and serum, were collected from each rat at 0.5 h, 1.0 h, and 2.0 h after oral administration of ZSS aqueous extract (15 g·kg-1). All calibration curves exhibited good linearity (r > 0.994 6) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at four different levels were both less than 19.77%, and the accuracies (RE) ranged from -19.68% to 19.46%; The extraction recoveries of the eight components ranged from 86.70% to 114.29%, and the matrix effects were from 82.14% to 114.57%. The validated method was successfully applied to the tissue distribution study of the eight components. The levels of swertisin, spinosin, 6‴-feruloylspinosin, and kaempferol-3-O-rutinoside in the small intestine were highest, then followed by the kidney, heart, and liver. Meanwhile, the levels of jujuboside A (JuA), jujuboside B (JuB), and jujuboside A1 (JuA1) in the small intestine were highest, then followed by the lung, spleen, and kidney. The concentrations of betulinic acid in the small intestine were higher than heart, lung, kidney, and liver. The flavonoids and saponins of ZSS with extremely low content could pass through the blood-brain barrier. The research results will provide an experimental basis for explaining the mechanism of nourishing the heart and tranquilizing the mind of ZSS. The animal experimental operations involved in this study followed the regulations of the Animal Ethics Committee of Shanxi University of Chinese Medicine and passed the animal experimental ethical review (No. 2021DW172).

16.
Journal of China Pharmaceutical University ; (6): 77-88, 2023.
Artículo en Chino | WPRIM | ID: wpr-965257

RESUMEN

@#Heibu Yaogao (ointment), which made of Galla chinensis, Scolopendra, etc., is a commonly used hospital preparation with definite clinical efficacy, yet its material basis has not been fully clarified.By ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), the chemical constituents of Heibu Yaogao (ointment) and its crude drug mixture were qualitatively analyzed, and the differences of components before and after preparation were discussed.Scanned by electrospray ionization (ESI) source in positive and negative ion modes, a total of 73 compounds were identified via precise molecular weight, characteristic fragment ions and chromatographic retention time.57 and 65 compounds were detected in crude drug mixture and Heibu Yaogao (ointment) respectively, including tannins, phenolic acids, nucleosides, amino acids, etc.In the process of preparation, the chemical structure changes are mainly concentrated in tannins.The change of components of Heibu Yaogao (ointment) before and after processing were proved by UPLC-Q-TOF-MS/MS, which provides important reference for elucidating the material basis of Heibu Yaogao (ointment), illustrating the rationality of preparation technology and further promoting the quality control of the preparation.

17.
Chinese Pharmacological Bulletin ; (12): 357-366, 2023.
Artículo en Chino | WPRIM | ID: wpr-1013864

RESUMEN

Aim To explore the mechanism of process¬ing and increasing efficiency of Arisaematis rhizomz preparatum. Methods UPLC-Q-TOF-MS/MS tech¬nology was used to detect the chemical components be¬fore and after processing of Arisaematis rhizomz prepara¬tum, and its mechanism of action was analysed in the treatment of 44 asthma and phlegm " by using network pharmacology. A rat model of allergic asthma was es- tablished to compare the efficacy of Arisaematis rliizoma before and after processing. Results A total of 27 chemical components were identified, among which cur- cumin ,6-gingerol and other components increased after processing. Combined with the database prediction, the action mechanism of the 36 chemical components in the treatment of 44 asthma and phlegm" diseases was dis¬cussed and predicted through network pharmacology. The results of animal experiments showed that the effect of processed Arisaematis rhizoma on allergic asth¬ma was better than that of Arisaematis rhizoma, but there was no significant difference. Conclusions The addition of curcumin, 6-gingerol, camphor, demethyl- curcumin and other components after the processed Ari¬saematis rhizomz preparatum may be the reason for the synergistic effect of Arisaematis rhizomz preparatum in the treatment of allergic asthma.

18.
Chinese Pharmacological Bulletin ; (12): 489-497, 2023.
Artículo en Chino | WPRIM | ID: wpr-1013832

RESUMEN

Aim To prepare the sea cucumber enzy¬molysis fermentation liquid (SCEFL) by enzymatic hydrolysis of protease and fermentation of probiotics and to investigate the effect of SCEFL on the immunosup-pression induced by cyclophosphamide in mice and to explore its mechanism by metabomic method. Methods The immunosuppressive model was induced by in-traperitoneal injection of cyclophosphamide. C57BL/6J mice were randomly divided into normal group, model group, Levamisole group, SCEFL groups (at low, medium and high doses). The pathological changes of spleen were observed by HE staining. The proportion of CD4

19.
Chinese Pharmacological Bulletin ; (12): 1303-1311, 2023.
Artículo en Chino | WPRIM | ID: wpr-1013762

RESUMEN

Aim To explore the effects of isovitexin (IVT) on alcoholic fatty liver disease (AFLD) and its mechanism based on metabolomics and in vivo methods and combined molecular docking. Methods 8-week-old male C57BL/6J mice were randomly divided into control, model and IVT groups, with 6 mice in each group. The control group was fed with alcoholic liquid feed control feed, the model group and IVT group were fed with alcoholic liquid feed model feed, and the IVT group was fed daily gastric IVT (100 mg • kg

20.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 140-150, 2023.
Artículo en Chino | WPRIM | ID: wpr-973755

RESUMEN

ObjectiveTo study the potential quality marker (Q-marker) of Tinosporae Radix associated with efficacy of "relieving sore throat" based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), multivariate statistical analysis (MSA), and network pharmacology. MethodUPLC-Q-TOF-MS was used to identify the main chemical components in 18 batches of Tinosporae Radix. On this basis, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were employed to screen out the main marker components that caused differences between groups. Moreover, network pharmacology technology was applied to predict the potential "sore throat-relieving" components, and the molecular docking between the common components resulting from MSA and network pharmacology and the core targets was carried out to verify the marker components. ResultA total of 17 compounds, including alkaloids, diterpenoid lactones, and sterols, were identified by UPLC-Q-TOF-MS. Five main differential components were found by MSA: Columbamine, jatrorrhizine, palmatine, menisperine, and columbin. Network pharmacology analysis yielded six compounds: tetrahydropalmatine, palmatine, menisperine, fibleucin, neoechinulin A, and columbin which were selected as potential "sore throat-relieving" components of Tinosporae Radix. They may relieve sore throat by acting on interleukin-6, epidermal growth factor receptor, prostaglandin G/H synthase 2, matrix metalloproteinase-9, proto-oncogene tyrosine-protein kinase Src and other targets, and regulating Hepatitis B, influenza A, human T-cell virus infection, human cytomegalovirus infection, coronavirus disease-2019, and other signaling pathways. The common active components in Tinosporae Radix resulting from MSA and network pharmacology analysis were palmatine, menisperine, and columbin, which had high binding affinity with six core targets and can be used as the Q-marker components of Tinosporae Radix in "relieving sore throat". ConclusionThis study predicts the "sore throat-relieving" Q-marker of Tinosporae Radix, which lays a basis for developing the quality standard of Tinosporae Radix based on the efficacy and improving the quality evaluation system of the medicinal.

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