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Prunella vulgaris L. (P. vulgaris) has great application value and development prospects in improving sleep. In this study, we continued to evaluate the sleep-improvement function and mechanism of P. vulgaris from both chemical characterization and function based on sleep-improvement functional ingredients, rosmarinic acid and salviaflaside, screened out in the previous stage as the index components. The chemical constituents of P. vulgaris and its phenolic acid fraction were characterized by the UPLC-MSn technology. The quality of the sleep-improvement phenolic acid fraction of P. vulgaris was scientifically evaluated by fingerprints combined with quantitative analysis of rosmarinic acid and salviaflaside. The function of phenolic acid parts of P. vulgaris in improving sleep was verified by different insomnia models including the PCPA-induced insomnia model and surface platform sleep deprivation model. HE staining was used to observe the effect of P. vulgaris on the morphology of nerve cells in different brain regions. In vivo experiments and molecular docking explored the sedative-hypnotic effects of functional ingredients of P. vulgaris. All these results investigated the material basis and mechanism of P. vulgaris to improve sleep from multiple perspectives, which contribute to providing a basis for the development of functional food to improve sleep.
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Depsidos , Extractos Vegetales , Prunella , Ácido Rosmarínico , Sueño , Prunella/química , Animales , Sueño/efectos de los fármacos , Depsidos/análisis , Extractos Vegetales/farmacología , Extractos Vegetales/química , Masculino , Cinamatos/análisis , Simulación del Acoplamiento Molecular , Trastornos del Inicio y del Mantenimiento del Sueño/tratamiento farmacológico , Hidroxibenzoatos/análisis , Ratones , Hipnóticos y Sedantes/farmacologíaRESUMEN
To fully understand whether Saposhnikoviae Radix polysaccharides(SP) can be metabolized in gastric fluid and the meta-bolic behavior, this study systematically analyzed the metabolites in simulated gastric fluid of SP by high-performance liquid chromatography-ion trap time-of-flight mass spectrometry(HPLC-IT-TOF-MS) technology in combination with zebrafish immune activity evaluation. Based on the obtained accurate relative molecular mass, chromatographic retention behavior, MS fragmentation patterns, refe-rence standards, and relevant literature reports, 19 metabolites were analyzed and identified. Among them, five monosaccharides and 14 oligosaccharides were generated as metabolites. Several reducing sugars, including mannose, glucose, rhamnose, and xylose, were accurately identified in the gastric fluid metabolites. Zebrafish pharmacological evaluation results indicated that SP maintained good immune activity after gastric fluid metabolism, with the most significant increase in immune cell density observed at W3(simulated gastric fluid metabolism for 2 hours). Among the gastric fluid metabolites, M1 and M3(Hex-Hex-Man) may be most closely related to pharmacological activity and could be further studied as potential active fragments.
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Medicamentos Herbarios Chinos , Pez Cebra , Humanos , Animales , Cromatografía Líquida de Alta Presión/métodos , Raíces de Plantas/química , Polisacáridos/análisis , Medicamentos Herbarios Chinos/químicaRESUMEN
The immunomodulatory effect of Saposhnikoviae Radix polysaccharide(SRP) was evaluated based on the zebrafish mo-del, and its mechanism was explored by transcriptome sequencing and real-time fluorescence-based quantitative PCR(RT-qPCR). The immune-compromised model was induced by navelbine in the immunofluorescence-labeled transgenic zebrafish Tg(lyz: DsRed), and the effect of SRP on the density and distribution of macrophages in zebrafish was evaluated. The effect of SRP on the numbers of macrophages and neutrophils in wild-type AB zebrafish was detected by neutral red and Sudan black B staining. The content of NO in zebrafish was detected by DAF-FM DA fluorescence probe. The content of IL-1ß and IL-6 in zebrafish was detected by ELISA. The differentially expressed genes(DEGs) of zebrafish in the blank control group, the model group, and the SRP treatment group were analyzed by transcriptome sequencing. The immune regulation mechanism was analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment, and the expression levels of key genes were verified by RT-qPCR. The results showed that SRP could significantly increase the density of immune cells in zebrafish, increase the number of macrophages and neutrophils, and reduce the content of NO, IL-1ß, and IL-6 in immune-compromised zebrafish. The results of transcriptome sequencing analysis showed that SRP could affect the expression level of immune-related genes on Toll-like receptor pathway and herpes simplex infection pathway to affect the release of downstream cytokines and interferon, thereby completing the activation process of T cells and playing a role in regulating the immune activity of the body.
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Interleucina-6 , Pez Cebra , Animales , Pez Cebra/genética , Interleucina-6/genética , Perfilación de la Expresión Génica , Citocinas/genética , Macrófagos , TranscriptomaRESUMEN
The combination of vascular endothelial growth factor (VEGF) inhibitors and tyrosine kinase inhibitors (TKIs) is newly available for molecular targeted therapy against non-small cell lung cancer (NSCLC) in clinic. However, the therapeutic benefits remain unsatisfying due to the poor drug delivery to targets of interest. In this study, we developed bevacizumab-coated gefitinib-loaded nanoparticles (BCGN) with dual-responsive drug release for inhibiting tumor angiogenesis and phosphorylation of epidermal growth factor receptor (EGFR). Through an exogenous corona strategy, bevacizumab is easily coated on gefitinib-loaded nanoparticles via electrostatic interaction. After intravenous injection, BCGN are efficiently accumulated in NSCLC tumors as confirmed by dual-model imaging. Bevacizumab is released from BCGN upon oxidation in tumor microenvironment, whereas gefitinib is released after being internalized by tumor cells and disassembled in reduction cytoplasm. The dual-responsive release of bevacizumab and gefitinib significantly inhibits tumor growth in both A549 and HCC827 human NSCLC models. Our approach provides a promising strategy to improve combinational molecular targeted therapy of NSCLC with precisely controlled drug release.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Humanos , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Gefitinib , Bevacizumab/uso terapéutico , Neoplasias Pulmonares/patología , Factor A de Crecimiento Endotelial Vascular , Terapia Molecular Dirigida , Quinazolinas/farmacología , Resistencia a Antineoplásicos , Línea Celular Tumoral , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Microambiente TumoralRESUMEN
BACKGROUND: Early thrombolytic therapy is crucial to treat acute cerebral infarction, especially since the onset of thrombolytic therapy takes 1-6 h. Therefore, early diagnosis and evaluation of cerebral infarction is important. AIM: To investigate the diagnostic value of magnetic resonance multi-delay three-dimensional arterial spin labeling (3DASL) and diffusion kurtosis imaging (DKI) in evaluating the perfusion and infarct area size in patients with acute cerebral ischemia. METHODS: Eighty-four patients who experienced acute cerebral ischemia from March 2019 to February 2021 were included. All patients in the acute stage underwent magnetic resonance-based examination, and the data were processed by the system's own software. The apparent diffusion coefficient (ADC), average diffusion coefficient (MD), axial diffusion (AD), radial diffusion (RD), average kurtosis (MK), radial kurtosis (fairly RK), axial kurtosis (AK), and perfusion parameters post-labeling delays (PLD) in the focal area and its corresponding area were compared. The correlation between the lesion area of cerebral infarction under MK and MD and T2-weighted imaging (T2WI) was analyzed. RESULTS: The DKI parameters of focal and control areas in the study subjects were compared. The ADC, MD, AD, and RD values in the lesion area were significantly lower than those in the control area. The MK, RK, and AK values in the lesion area were significantly higher than those in the control area. The MK/MD value in the infarct lesions was used to determine the matching situation. MK/MD < 5 mm was considered matching and MK/MD ≥ 5 mm was considered mismatching. PLD1.5s and PLD2.5s perfusion parameters in the central, peripheral, and control areas of the infarct lesions in MK/MD-matched and -unmatched patients were not significantly different. PLD1.5s and PLD2.5s perfusion parameter values in the central area of the infarct lesions in MK/MD-matched and -unmatched patients were significantly lower than those in peripheral and control areas. The MK and MD maps showed a lesion area of 20.08 ± 5.74 cm2 and 22.09 ± 5.58 cm2, respectively. T2WI showed a lesion area of 19.76 ± 5.02 cm2. There were no significant differences in the cerebral infarction lesion areas measured using the three methods. MK, MD, and T2WI showed a good correlation. CONCLUSION: DKI parameters showed significant difference between the focal and control areas in patients with acute ischemic cerebral infarction. 3DASL can effectively determine the changes in perfusion levels in the lesion area. There was a high correlation between the area of the infarct lesions diagnosed by DKI and T2WI.
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The calibration of chromone reference extract(CRE) was conducted and a quality control method of Saposhnikoviae Radix(SR) was established based on CRE. Meanwhile, the quality control system of SR was improved and the feasibility of using reference extract as a substitute for single reference substance in quality control of Chinese medicine was discussed. In this study, the content of the prepared CRE was calibrated with prim-O-glucosylcimifugin, cimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, and secO-glucosylhamaudol as indicators. Subsequently, an HPLC analytical method was developed to determine the content of four chromones in 20 batches of SR samples based on the CRE with known content as the standard substance. T-test was used for the comparison of the determination results of the two methods(single chemical component and CRE as reference substances, respectively), and the P values of prim-O-glucosylcimifugin, cimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, and sec-O-glucosylhamaudol were 0. 16,0. 39, 0. 14, and 0. 42. The results demonstrated that there was no significant difference between the two methods. This study initially verified the feasibility that the CRE could be used as a substitute for single reference substance in quality control of SR. In conclusion,this study is expected to provide a scientific basis and a new research model for the application of reference extract in the quality control of Chinese medicine.
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Apiaceae , Medicamentos Herbarios Chinos , Calibración , Cromatografía Líquida de Alta Presión , Cromonas , Control de CalidadRESUMEN
We aimed to assess the risks of Cryptosporidium and Giardia infections associated with drinking water for local residents, based on a quantitative microbial risk assessment, in three densely populated regions of China. In total, 45 source water samples and 45 treated water samples were collected from June to December 2014. Five Cryptosporidium-positive samples and 5 Giardia-positive samples were found. The annual probability of infection for individuals in Jintan (6.27 × 10 -4-2.05 × 10 -3 for Cryptosporidium and 7.18 × 10 -4-2.32 × 10 -3 for Giardia), Ezhou (6.27 × 10 -4-1.10 × 10 -2 for Cryptosporidium and 3.65 × 10 -4-1.20 × 10 -3 for Giardia), and Binyang (3.79 × 10 -4-1.25 × 10 -3 for Cryptosporidium) exceeded the tolerable risk of infection of 10 -4 set by the United States Environmental Protection Agency. Moreover, the corresponding disease burdens of cryptosporidiosis and giardiasis, due to direct drinking and residual water in these regions, exceeded the threshold of 10 -6 disability-adjusted life years per person per year set by the World Health Organization. These results provide insights into strategies to improve the safety of drinking water.
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Cryptosporidium/aislamiento & purificación , Giardia/aislamiento & purificación , Microbiología del Agua , Abastecimiento de Agua/estadística & datos numéricos , China , Criptosporidiosis/microbiología , Giardiasis/microbiología , Humanos , Medición de RiesgoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Prunella vulgaris L. (P. vulgaris) is a medicinal plant belonging to the Labiatae family, and its dried spikes is called as Xiakucao in China, which is a common traditional Chinese medicine with the activities of clearing the liver and expelling fire, improving eyesight, dispersing nodules and detumescence. Modern pharmacological studies have proved that P. vulgaris has various pharmacological activities such as immunomodulatory, antiviral, antibacterial and anti-insomnia activities. AIMS OF THIS REVIEW: P. vulgaris have been reported to have anti-insomnia effects. Nevertheless, the pharmacodynamic substance basis of this anti-insomnia effect is still unclear. The aim of this study was to identify the active components responsible for evoking the anti-insomnia effect of P. vulgaris and to evaluate its anti-insomnia effect. MATERIALS AND METHODS: In this study, we proposed a method combined with pharmacodynamic experiments, extraction and enrichment of chemical components, and the plasma pharmacochemistry to screen out the anti-insomnia components of P. vulgaris. Firstly, the active eluted fraction of the ethanol extract was screened out based on pharmacodynamic tracing method, and then the chemical composition was analyzed systematically by UPLC-MS/MS. Thirdly, pharmacodynamic tracing method and silica gel column chromatography were employed to screen out the active fraction of 70% ethanol eluted fraction, and its bioactive components in vitro and in vivo were identified by UPLC-MS/MS. Finally, screening out the anti-insomnia components of P. vulgaris by comparing the difference between in vivo and in vitro components, and three potentially bioactive ingredients were validated experimentally. RESULTS: It was confirmed that the fraction eluted with 70% ethanol from macroporous adsorption resin column was responsible for the anti-insomnia efficacy, and 55 compounds were identified or preliminarily identified. Then totally 9 compounds in vitro and 12 compounds in vivo from the active fraction of 70% ethanol eluted fraction were tentatively identified. Among them, mangiferin, rosmarinic acid and salviaflaside were the prototype components of P. vulgaris, which indicated that the three compounds might play the key role in the anti-insomnia activities. In vivo, compared to blank control group, the three compounds significantly shortened the sleeping latency and prolonged the sleeping time produced by pentobarbital sodium. CONCLUSIONS: This study clarified that mangiferin, rosmarinic acid and salviaflaside were considered as the anti-insomnia components of P. vulgaris. This is the first study on screening out the active ingredients responsible for evoking the anti-insomnia effect of P. vulgaris. The three compounds of P. vulgaris may help develop one or more drugs to prevent or treat insomnia. Further investigations are recommended to define the mechanism of the anti-insomnia activity of P. vulgaris.
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Extractos Vegetales/farmacología , Prunella/química , Trastornos del Inicio y del Mantenimiento del Sueño/tratamiento farmacológico , Animales , Cromatografía Líquida de Alta Presión , Cinamatos/aislamiento & purificación , Cinamatos/farmacología , Depsidos/aislamiento & purificación , Depsidos/farmacología , Glucósidos/aislamiento & purificación , Glucósidos/farmacología , Masculino , Ratones , Ratones Endogámicos ICR , Fenilpropionatos/aislamiento & purificación , Fenilpropionatos/farmacología , Extractos Vegetales/química , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem , Xantonas/aislamiento & purificación , Xantonas/farmacología , Ácido RosmarínicoRESUMEN
By establishing the preparation process of Scrophulariaceae Radix reference extract(SRRE) and calibrating it, we discussed its feasibility as a substitute for single reference substance in the quality control of Scrophulariae Radix. The SRREs were prepared by solvent extraction method and chromatographic separation technology, and then calibrated with the reference substances of harpagide, angoroside C and harpagoside. The HPLC content determination method of Scrophulariae Radixl was established with SRREs of the known content and the reference substances of harpagide, angoroside C and harpagoside respectively as the control ones. Then the content of three components in Scrophulariae Radix was determined, and the t-test method was used to compare the results of the two methods. With SRRE as references, harpagide, angoroside C and harpagoside were in a good linear relationship(r≥0.999 8) within each range, and the average recovery rate was 98.55% to 100.6%. The t-test results showed that the P values of two determination methods were 0.493, 0.155 and 0.171 for harpagide, angoroside C and harpagoside respectively, indicating no significant diffe-rence between the two methods of content determination. The SRRE can be used as a substitute for the reference in the quality control of Scrophulariaceae Radix. The SRRE can replace the corresponding reference substance for the quality control of Scrophulariae Radix. The results of this study provide new methods and new ideas for the quality evaluation of Scrophulariae Radix, and provide a scientific basis for the application of reference extracts in the quality research of traditional Chinese medicine.
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Medicamentos Herbarios Chinos , Scrophularia , Scrophulariaceae , Cromatografía Líquida de Alta Presión , Control de CalidadRESUMEN
Chinese traditional medicine compound is the main form of Chinese medicine clinical application. The elucidation of the effective components of traditional Chinese medicine is one of the key scientific issues to promote the modernization of traditional Chinese medicine. At present, there are many research ideas on the effective components of traditional Chinese medicine compounds. By analyzing the current status and existing problems of existing research ideas, the author proposes a "double reduction network pharmacology"(2 R network pharmacology) research method based on "prediction of dominant components-potential target selection". Chemical components with good properties were selected by ADMET property prediction technology, and compared with the blood components and target organ components to determine the dominant components with potential therapeutic effect, that is "reducing constituents"; the potential core regulatory pathway of traditional Chinese medicine compound was enriched by RNA-Seq technology combined with network database, and then the target of traditional Chinese medicine compound was mined based on the signal pathway, that is "reducing targets". To improve the efficiency and accuracy of effective component screening, the network relationship of "component target" was established by the related technology of network pharmacology. The purpose of this study is to provide practical research ideas and methods for clarifying the effective components of traditional Chinese medicine, revealing the law of compatibility of traditional Chinese medicine and clarifying the target of drug action.
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Medicamentos Herbarios Chinos , Medicina Tradicional China , Bases de Datos Factuales , Medicamentos Herbarios Chinos/farmacología , Simulación del Acoplamiento Molecular , Proyectos de InvestigaciónRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Clematis chinensis Osbeck (C. chinensis), Clematis hexapetala Pall (C. hexapetala) and Clematis terniflora var. mandshurica Rupr (C. mandshurica) are collectively referred to as Clematidis Radix et Rhizome (CRR) in China. CRR is widely distributed in China, which is used as a traditional Chinese medicine to treat rheumatic arthralgia, limb numbness, tendon constriction and inconvenience in flexion and extension. AIMS OF THIS REVIEW: This review systematically summarized the research progress on uses, chemical components, pharmacological activities and toxicology of CRR, listed the chemical structures of main compounds for clarifying the differences in chemical compositions. Meanwhile, the review will provide a theoretical and practical basis for the further research and development of CRR. MATERIALS AND METHODS: The available information on CRR was collected using published materials and electronic databases, including ancient and modern books, Chinese Pharmacopoeia, Ph.D. and M. Sc. dissertations, CNKI, SciFinder, WanFang data, PubMed, ScienceDirect and Web of Science. The starting and ending years of references is 1965-2020, the search strategy was conducted by key words such as uses, chemical components, pharmacology and toxicology of CRR. RESULTS: Up to now, CRR has been used to treat various diseases/disorders, such as relieving rheumatism pain, treating cervical spondylopathy and scapulohumeral periarthritis, treating hepatic carcinoma and gastrointestinal, etc. In addition, more than 200 compounds have been isolated from the three plant species of Clematidis. Moreover, the crude extracts and isolated compounds of CRR have been reported to have a wide range of pharmacological activities, such as anti-inflammatory, anti-tumor, antimicrobial and antioxidant activities, etc. Toxicity studies have shown that CRR can cause oral burning, swelling, abdominal pain or severe diarrhea, difficulty breathing, dilated pupils, renal tissue structural changes, and severe death. CONCLUSIONS: Researches in recent years mainly focused on C. chinensis and C. mandshurica, while there are a few reports on the pharmacological studies of C. hexapetala. Therefore, it is necessary to conduct further research on C. hexapetala. Meanwhile, it is important to pay attention to pursue research on the similarities and differences between the three plant species of Clematidis to find their respective advantages and make rational use of CRR. In addition, there is no report on the mechanism of toxicity research, which needs more attention.
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Clematis/química , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Fitoquímicos/química , Fitoquímicos/farmacología , Raíces de Plantas/química , Rizoma/química , Animales , Medicamentos Herbarios Chinos/uso terapéutico , Medicamentos Herbarios Chinos/toxicidad , Humanos , Fitoquímicos/uso terapéutico , Fitoquímicos/toxicidadRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Si-Miao-Yong-An decoction (SMYAD), a classical traditional Chinese medicine (TCM) formula, has been used to treat various cardiovascular diseases in clinics. AIM OF THE STUDY: The aim of this study is to investigate the effective combinatorial components from SMYAD and its mechanism regarding the intervention on myocardial hypertrophy. MATERIALS AND METHODS: SMYAD constituents absorbed in rat plasma and heart were identified using UHPLC Q-Exactive-Orbitrap MS/MS. The identified constituents in SMYAD were further analyzed using ADMET (absorption, distribution, metabolism, excretion and toxicity) prediction and molecular docking. The effective constituents were identified using isoproterenol (ISO)-induced H9c2 cardiomyocyte hypertrophy, and neochlorogenic acid (NCA), chlorogenic acid (CA), cryptochlorogenic acid (CCA), isochlorogenic acid C (ICAC), angoroside C (AGDC), isochlorogenic acid A (ICAA), sweroside (SRD), and harpagide (HPD) in SMYAD extract were quantified by HPLC for compatibility. Finally, anti-hypertrophic activities of candidate effective combinatorial components, which were prepared according to the determined molar concentration ratio of effective constituents using reference substance solution, were analyzed using immunofluorescence staining and Quantitative real-time PCR. The expression levels of PI3Kα, p-ERK, p-Akt, Akt, p-mTOR, mTOR and HIF-1α were measured using Western blot. RESULTS: 32 prototypes of SMYAD were identified from plasma and heart tissue of rat. Combining with ADMET prediction, 31 dominant constituents were focused. Based on HIF-1 pathway identified in preliminary result, 17 targets were focused, which were used to dock with 31 constituents. 27 constituents were therefore hit as the potential effective constituents of SMYAD in inhibiting myocardial hypertrophy. Bioactivity evaluation showed that NCA, CA, CCA, ICAC, AGDC, ICAA, SRD, and HPD significantly inhibited the increase of H9c2 cell surface area induced by ISO. Except for ICAA and AGDC, the remaining 6 effective constituents, showing a certain inhibitory effect on ISO-induced ANP mRNA overexpression at high and low concentrations, participated in compatibility based on the molar concentration ratio determined by HPLC. Effective combinatorial components composed of the 6 effective constituents (effective combinatorial components ABC) showed significant inhibitory effect on the increase of cell surface area, and the overexpression of ANP and ß-MHC mRNA in H9c2 cells induced by ISO. Moreover, effective combinatorial components ABC significantly inhibited the protein overexpressions of p-Akt, p-mTOR and HIF-1α. Based on the results, we put forward the strategy of "Focusing constituents" and "Focusing targets" for the effective constituents research of TCM formula. CONCLUSION: Effective combinatorial components ABC composed of NCA, CA, CCA, ICAC, SRD and HPD from SMYAD inhibited ISO-induced cardiomyocyte hypertrophy and down-regulated expression of ANP and ß-MHC mRNA through the inactivation of Akt/mTOR/HIF-1α pathway.
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Cardiomegalia/tratamiento farmacológico , Cardiomegalia/metabolismo , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Animales , Factor Natriurético Atrial/genética , Línea Celular , Medicamentos Herbarios Chinos/metabolismo , Medicamentos Herbarios Chinos/uso terapéutico , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Isoproterenol/toxicidad , Masculino , Medicina Tradicional China , Simulación del Acoplamiento Molecular , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Cadenas Pesadas de Miosina/genética , Fosfatidilinositol 3-Quinasa/metabolismo , Fitoquímicos/análisis , Fitoquímicos/farmacología , Fitoquímicos/uso terapéutico , Plasma/química , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas Sprague-Dawley , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
Current chemotherapy regimens on acute myeloid leukemia (AML) still have some drawbacks, such as intolerance and drug resistance, which calls need for the development of targeted therapy. Signal transducer and activator of transcription 5 (STAT5) is often overexpressed or abnormally activated in leukemia and involved in cell self-renewal, proliferation, and stress adaptation. Overexpressed Aurora A (AURKA) is associated with poor prognosis in tumors, and inhibitors against AURKA are already in clinical trials. However, it has rarely been reported whether AURKA inhibitors restrain STAT5-activated leukemia cells. In this study, we constructed STAT5 constitutively activated (cS5) cells and found that STAT5 promoted cell proliferation and colony formation. Moreover, cS5 cells showed elevated reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels, which indicated higher mitochondrial metabolism in cS5 cells. A novel AURKA inhibitor AKI604 was synthesized and showed significant inhibitory effects to the proliferation and colony formation in both STAT5 constitutively activated and nonactivated AML cells. AKI604 induced mitochondrial impairment, leading to the disruption of mitochondrial membrane potential and the elevation of ROS as well as cellular calcium (Ca2+ ) levels. AKI604 could also decline basal oxygen consumption rate and ATP biosynthesis, indicating the damage of oxidative phosphorylation. Furthermore, AKI604 exhibited significant antitumor effect in the HL-60 cS5 xenograft model of the BALB/c nude mice without an obvious influence on mice body weight and other healthy indicators. This study suggested that AKI604 was a potential strategy to overcome STAT5-induced leukemic proliferation in AML treatment by inducing mitochondrial impairment.
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Antineoplásicos/farmacología , Aurora Quinasa A/antagonistas & inhibidores , Leucemia Mieloide Aguda/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Células HL-60 , Humanos , Leucemia Mieloide Aguda/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Factor de Transcripción STAT5/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The study on the targets of traditional Chinese medicine is an important part of researchers using modern scientific language to clarify the mechanism of traditional Chinese medicine. However, the research on the targets of Chinese medicine is full of challenges due to the complexity of active ingredients. As a branch of systems biology, proteomics focus on specific proteins in living organisms from a holistic perspective, which significantly improves the efficiency of targets discovery and has obvious advantages in the research of targets of Chinese medicine. Based on relevant literature and different methods used in targets of Chinese medicine, proteomics can be divided into chemical proteomics, differential proteomics and quantitative proteomics. The applications of the above three methods are illustrated in this paper as well, which will provide new methods and ideas for the study of the mechanism of Chinese medicine in the future.
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Medicamentos Herbarios Chinos/análisis , Medicina Tradicional China , ProteómicaRESUMEN
Flacourtiaceae plants are widely used as folk medicines in traditional medicine systems for its chemical diversity and pharmacological activities. In many different areas, Flacourtiaceae plants are used as traditional medicines for the treatment of ulcers, malaria, rheumatism. The Flacourtiaceae plants contain a very plentiful chemical composition, and phytochemical studies show that the Flacourtiaceae plants contained terpenoids, aromatic glycosides, flavnoids, phenylpropanoids, alkaloids, fatty hydrocarbon, and other compounds. In pharmacological studies, various extract and isolated individual compounds exhibited antitumor, anti-oxidation, and anti-inflammatory activities. In this review, the literature data on the chemical constituents and pharmacological investigations of the Flacourtiaceae plants are summarized, to provide information about a more comprehensive chemical composition and detailed pharmacological activities of Flacourtiaceae plants, with a view of further development of clinical medication. However, research on quantitative analysis, toxicity, and drug safety in vitro and in vivo is still insufficient, and further research is required.
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Fitoterapia , Extractos Vegetales/farmacología , Salicaceae/química , Antiinflamatorios , Antineoplásicos , Antioxidantes , Flavonoides/análisis , Glicósidos/análisis , Humanos , Malaria/tratamiento farmacológico , Fitoquímicos/análisis , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/uso terapéutico , Enfermedades Reumáticas/tratamiento farmacológico , Terpenos/análisis , Úlcera/tratamiento farmacológicoRESUMEN
Menthae Haplocalycis Herba has been utilized for food and medicinal purposes in China for thousands of years. It has various efficacies, including dispelling wind and heat and relieving sore throat. M. Haplocalycis Herba has been also widely used in food, cosmetics, spices, and other fields. Exploring the constituents and detecting the metabolites of M. Haplocalycis are of great significance to clarify the effective substances. However, the in vivo metabolites of M. Haplocalycis Herba water extract are still unclear. Herein, a sensitive and specific method, ultra-high performance liquid chromatography with linear ion trap-Orbitrap mass spectrometry, established in this assay was used to study the metabolism of M. Haplocalycis Herba water extract in rat plasma, urine, and feces. We characterized and identified 9, 50, and 34 metabolites in plasma, urine, and feces, respectively. Seven metabolic pathways, including phase â (isomerization, demethylation, hydroxylation, and dehydration) and phase â ¡ (sulfation and glucuronidation) were mainly involved in the metabolism. It is the first systematic study on the metabolism of M. Haplocalycis Herba water extract in vivo, which enrich current understanding of the metabolic behavior of M. Haplocalycis Herba water extract and provide a metabolic rationale for further in-depth in vivo biotransformation and pharmacokinetic analysis.
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Medicamentos Herbarios Chinos/análisis , Heces/química , Mentha/química , Fitoquímicos/análisis , Extractos Vegetales/análisis , Animales , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/metabolismo , Inyecciones Intravenosas , Masculino , Espectrometría de Masas , Medicina Tradicional China , Mentha/metabolismo , Estructura Molecular , Fitoquímicos/administración & dosificación , Fitoquímicos/metabolismo , Extractos Vegetales/administración & dosificación , Extractos Vegetales/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
This experiment was performed to analyze and identify the chemical constituents of Lycii Cortex by UPLC-LTQ-OrbitrapMS. The analysis was performed on a Waters Xbridge Shield RP18( 4. 6 mm×250 mm,5 µm) column with the mobile phase of 0. 1%formic acid( A)-acetonitrile( B) under gradient conditions at a flow rate of 1. 0 m L·min-1 and the temperature maintained at 35 â ï¼Electrospray ionization ion trap time-off light multistage mass spectrometry was applied for qualitative analysis under positive and negative ion modes. The results indicated that 55 compounds consisted of 39 phenolic amides,6 organic acids,3 cyclic peptides,2 coumarins and 5 others. In conclusion,an UPLC-LTQ-Orbitrap-MS method was established to qualitative analysis of Lycii Cortex in this study,and the fragmentation rules of phenolic amides were summarized,which provides a good foundation for further study of Lycii Cortex.
Asunto(s)
Medicamentos Herbarios Chinos/química , Cromatografía Líquida de Alta Presión , Cumarinas , Espectrometría de Masas , FenolesRESUMEN
The tea-like beverage Stevia rebaudiana Bertoni (Stevia) is popular in China because it reduces blood glucose and has a sweet taste. In this work, a comprehensive quality assessment of Stevia led to the discovery of five phenylethanoid glycosides, namely steviophethanoside (1), cuchiloside (2), salidroside (3), icariside D (4), and tyrosol (5). Of them, compound 1 is a novel compound. Mass spectrometry and NMR spectroscopy were employed to confirm the absolute configuration. A hydrolytic step with 4 N TFA at 95 °C for 4 h was used to confirm the monosaccharides. In addition, Discovery Studio 4.0 was used to predict the ADME and toxicity activity of compound 1. The results suggested that compound 1 was biocompatible and had poor toxicity, which was verified by rat INS-1 islet ß cells through an MTT assay. Meanwhile, a significant stimulatory effect on INS-1 cells was observed, which indicated a hypoglycemic effect of compound 1. This is the first report that describes a natural, novel, and hypoglycemic phenylethanoid glycoside in Stevia.
Asunto(s)
Glicósidos/farmacología , Secreción de Insulina/efectos de los fármacos , Células Secretoras de Insulina/efectos de los fármacos , Extractos Vegetales/farmacología , Stevia/química , Animales , Células Cultivadas , China , Glicósidos/química , Glicósidos/aislamiento & purificación , Células Secretoras de Insulina/metabolismo , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , RatasRESUMEN
The main chemical constituents of naphthopyrone reference extract( NRE) with definite content and relatively fixed chemical composition were analyzed and determined. Ultra-high performance liquid chromatography-LTQ-Orbitrap XL mass spectrometry and high performance liquid chromatography were used to systematically study NRE from the aspects of main chemical components and determination. The results showed that the chemical composition of naphthopyrone reference extract of Cassiae Semen was relatively fixed,and seven naphthalopyranones were identified. Cassiaside B_2,cassiaside C_2,rubrofusarin-6-O-ß-D-gentiobioside and cassiaside C were the main chemical constituents of NRE,of which the determination and uncertainty results were( 11. 40+ 0. 26) %,( 11. 68+0. 24) %,( 16. 60+0. 22) %,( 28. 8+0. 48) %,respectively. This study contributed to the accurate evaluation of NRE and the foundation for the application of NRE in the quality control of Cassiae Semen,and provided a new idea for the replacement of single chemical reference substance by the reference extract of traditional Chinese medicine.