RESUMEN
DT-13 combined with topotecan (TPT) showed stronger antitumour effects in mice subcutaneous xenograft model compared with their individual effects in our previous research. Here, we further observed the synergistically effect in mice orthotopic xenograft model. Metabolomics analysis showed DT-13 combined with TPT alleviated metabolic disorders induced by tumour and synergistically inhibited the activity of the aerobic glycolysis-related enzymes in vivo and in vitro. Mechanistic studies revealed that the combination treatment promoted epidermal growth factor receptor (EGFR) degradation through non-muscle myosin IIA (NM IIA)-induced endocytosis of EGFR, further inhibited the activity of hexokinase II (HK II), and eventually promoted the aerobic glycolysis inhibition activity more efficiently compared with TPT or DT-13 monotherapy. The combination therapy also inhibited the specific binding of HK II to mitochondria. When using the NM II inhibitor (-)002Dblebbistatin or MYH-9 shRNA, the synergistic inhibition effect of DT-13 and TPT on aerobic glycolysis was eliminated in BGC-823 cells. Immunohistochemical analysis revealed selective up-regulation of NM IIA while specific down-regulation of p-CREB, EGFR, and HK II by the combination therapy. Collectively, these findings suggested that this regimen has significant clinical implications, warranted further investigation.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma/tratamiento farmacológico , Glucólisis/efectos de los fármacos , Saponinas/uso terapéutico , Neoplasias Gástricas/tratamiento farmacológico , Topotecan/uso terapéutico , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Carcinoma/enzimología , Carcinoma/genética , Carcinoma/metabolismo , Línea Celular Tumoral , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Sinergismo Farmacológico , Endocitosis/efectos de los fármacos , Receptores ErbB/metabolismo , Femenino , Hexoquinasa/antagonistas & inhibidores , Hexoquinasa/metabolismo , Humanos , Espectroscopía de Resonancia Magnética , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Miosina Tipo IIA no Muscular/metabolismo , ARN Interferente Pequeño , Saponinas/farmacología , Neoplasias Gástricas/enzimología , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Topotecan/farmacología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Success in generating insulin-producing cells (IPCs) from human embryonic stem (hES) cells by genetic manipulation has recently revealed a new therapeutic potential for diabetes. However, clinical application has been hampered by the viral genome integration and the risk of insertion mutagenesis that are entailed. Herein, we report the induction of hEC into IPCs by direct delivery of human Pdx1 proteins per se. Recombinant human Pdx1 proteins (hPdx1), which have an Antennapedia-like protein transduction domain sequence in their structure, can be efficiently translocated into hES cells and function as pancreatic transcription factor. hPdx1 protein activates a group of genes related to pancreatic beta-cell lineage development in hES cells, including NeuroD1, Nkx2.2, Pax4, Pax6, Nkx6.1 and Isl-1. hPdx1-treated hES cells synthesise and release insulin in response to glucose challenge. Therefore, this study constitutes a proof-of-concept demonstration of protein-mediated pancreatic specific differentiation of the hES cells by exploiting specific intrinsic properties of the hPdx1 protein.
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Diferenciación Celular , Linaje de la Célula , Células Madre Embrionarias/citología , Proteínas de Homeodominio/metabolismo , Células Secretoras de Insulina/citología , Transactivadores/metabolismo , Línea Celular , Células Madre Embrionarias/metabolismo , Proteína Homeobox Nkx-2.2 , Humanos , Células Secretoras de Insulina/metabolismo , Proteínas Nucleares , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
OBJECTIVE: To establish the quality standard for Danmo capsule. METHODS: TLC was used for the qualitative identification of Salviae Miltiorrhizae Radix Et Rhizoma and Lonicerae Japonicae Flos. HPLC was used to determine the content of Salvianolic acid B. RESULT: TLC spots were clear and well-separated without negative interference. The linear range of Salvianolic acid B was 0.120042 - 2.40084 microg (r = 0.9999) with an average recovery of 103.63%, RSD was 0.6% (n = 6). CONCLUSION: The method is simple, accurate and reliable. It can be used for the quality control of Danmo capsule.
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Benzofuranos/análisis , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/normas , Lonicera/química , Salvia miltiorrhiza/química , Cápsulas/normas , Ácido Clorogénico/análisis , Combinación de Medicamentos , Medicamentos Herbarios Chinos/aislamiento & purificación , Etanol/química , Flores/química , Plantas Medicinales/química , Control de Calidad , Reproducibilidad de los Resultados , Rizoma/químicaRESUMEN
OBJECTIVE: To study the effects of modified Mahuang Fuzi Xixin Decoction extract (MFXDE) on ischemia/reperfusion induced atrioventricular (A-V) block in rabbits model. METHODS: New Zealand rabbits were randomly divided into six groups, three test groups (Group TA, TB, TC) and three control groups (Group CA, CB, CC), 10 in each. MFXDE 2 mL/kg was given twice a day by gastrogavage to the test groups, while to the control groups, equal volume of normal saline was given instead, for 3 successive days. Twenty minutes after the last gastrogavage, right coronary artery ligation was performed in rabbits under anesthesia with 1.2 g/kg of 20% urethane via ear marginal vein injection, and lasted for 15 min (Group TA and CA), 60 min (Group TB and CB), and 120 min (Group TC and CC), respectively. ECG lead-II and His Bundle ECG were recorded at different time points to observe P-R interval and A-H interval. RESULTS: P-R interval and A-H interval in the test groups were shorter than in the control groups significantly (all P < 0.05) at the time point of 5 min after ischemia; and at the reperfusion stage, a re-extending phenomenon of P-R and A-H could be found in the CC group after 60 min reperfusion (P < 0.05), but it didn't occure in all the other test groups. CONCLUSION: MFXDE could improve the ischemia/reperfusion induced A-V conductive function.
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Bloqueo Atrioventricular/tratamiento farmacológico , Medicamentos Herbarios Chinos/administración & dosificación , Daño por Reperfusión Miocárdica/complicaciones , Animales , Bloqueo Atrioventricular/etiología , Bloqueo Atrioventricular/fisiopatología , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Conejos , Distribución AleatoriaRESUMEN
OBJECTIVE: To provide scientific basis for the selection of agrotype and property fertilization for Rumex gmelini cultivated in compliance with good agricultural practice (GAP). METHOD: HPLC method was applied to determinate the content of seven active constituents (resveratrol, polydatin, chrysophanol 1-glucoside, nepodin, emodin, chrysophanol and physcion) of annual R. gmelini. And the correlation between soil nutrients and content of active constituents in the root of R. gmelini were analyzed by stepwise regression analysis. RESULT: Seven regression equation were established. The statistic significance was found in three of them. CONCLUSION: The soil with high total K level is not suitable for R. gmelini cultivation. But the higher available N, available P, available K level of soil is suitable.
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Raíces de Plantas/química , Plantas Medicinales/química , Rumex/química , Suelo/análisis , Antraquinonas/análisis , Ecosistema , Emodina/análogos & derivados , Emodina/análisis , Fertilizantes , Glucósidos/análisis , Nitrógeno/análisis , Fósforo/análisis , Raíces de Plantas/crecimiento & desarrollo , Plantas Medicinales/crecimiento & desarrollo , Potasio/análisis , Análisis de Regresión , Resveratrol , Rumex/crecimiento & desarrollo , Estilbenos/análisisRESUMEN
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that leads to severe multiorgan damage. Lang Chuang Fang (LCF) is a Chinese herbal medicine that is clinically prescribed for treating SLE. In this study, we examined the therapeutic effects of LCF granule on lupus-prone MRL/lpr mice. Female mice were randomly separated into six groups, and LCF treatment groups received LCF granule at the dosage of 0.97 g/kg/d, 1.95 g/kg/d, and 3.90 g/kg/d, respectively. Here, we found that, compared to the MRL/lpr mice, both the spleen coefficient and thymus coefficient were reduced in the LCF granule-treated mice. There was a marked downregulation in CRP and anti-dsDNA autoantibody and an evident upregulation of CH50 in LCF granule-treated mice. LCF granule treatment also obviously reduced the proteinuria, BUN, and SCr levels in MRL/lpr mice at the dosage of 0.97 g/kg/d, 1.95 g/kg/d, and 3.90 g/kg/d, indicating that LCF granule alleviated the renal injury of MRL/lpr mice. Furthermore, LCF granule decreased p65 NF-κB levels and increased Sirt1 and Nrf2 levels in the kidney tissues of MRL/lpr mice, which might elucidate the beneficial effects of LCF on lupus nephritis. In conclusion, this study demonstrates that LCF granule has therapeutic effects on lupus-prone MRL/lpr mice.
RESUMEN
Combination therapy has a higher success rate for many cancers compared to mono-therapy. The treatment of Topotecan (TPT) on gastric cancer (GC) is limited by its toxicity and the potential drug resistance. We found that the combination of the saponin monomer 13 from the dwarf lilyturf tuber (DT-13), performing anti-metastasis and anti-angiogenesis effects, with TPT synergistically induced apoptotic cytotoxicity in GCs with high EGF receptor (EGFR) expression, which was dependent on DT-13-induced endocytosis of EGFR. With TPT, DT-13 promoted EGFR ubiquitin--mediated degradation through myosin IIA-induced and Src/ caveolin-1 (Cav-1)-induced endocytosis of EGFR; inhibited EGFR downstream signalling and then increased the pro-apoptotic effects. Moreover, the synergistic pro-apoptotic efficacy of DT-13 and TPT in GCs with high EGFR expression was eliminated by both the NM II inhibitor (-)-blebbistatin and MYH-9 shRNA. The combination therapy of DT-13 with TPT showed stronger anti-tumour effects in vivo compared with their individual effects. Moreover, the results of combination therapy revealed selective upregulation of pro-apoptotic activity in TUNEL assays and cleaved caspase-3 and NM IIA in immunohischemical analysis; while specific downregulation of p-extracellular regulated kinase 1/2 (p-ERK1/2), EGFR and Cav-1 in immunohischemical analysis. Collectively, these findings have significant clinical implications for patients with tumours harbouring high EGFR expression due to the possible high sensitivity of this regimen.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Endocitosis/efectos de los fármacos , Receptores ErbB/metabolismo , Miosina Tipo IIA no Muscular/metabolismo , Saponinas/farmacología , Neoplasias Gástricas/tratamiento farmacológico , Topotecan/farmacología , Animales , Línea Celular Tumoral , Sinergismo Farmacológico , Femenino , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Saponinas/administración & dosificación , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Topotecan/administración & dosificación , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Cancer metastasis plays a major role in tumor deterioration. Metastatic processes are known to be regulated by hypoxic microenvironment and non-muscle myosin IIA (NMIIA). DT-13, a bioactive saponin monomer isolated from Ophiopogon japonicus, has been reported to inhibit various cancer metastasis, but whether NMIIA is involved in the anti-metastatic activity of DT-13 under hypoxia remains to be determined. Thus, this study aims to clarify the role of DT-13 in regulating 95D cell metastasis under hypoxic microenvironment and to further investigate whether NMIIA is involved in the anti-metastatic mechanism of DT-13. We found that DT-13 significantly inhibited 95D cells metastasis in vitro and in vivo. Furthermore, hypoxia significantly inhibited the expression of NMIIA and redistributed NMIIA to the cell periphery, whereas DT-13 reversed the hypoxic effects by upregulating the expression of NMIIA. Moreover, DT-13 treatment redistributed NMIIA to the nuclear periphery and reduced the formation of F-actin in 95D cells. In addition, we found that the Raf-ERK1/2 signaling pathway is involved in regulation of NMIIA by DT-13. Collectively, these findings support NMIIA as a target of DT-13 to prevent lung cancer metastasis.
Asunto(s)
Hipoxia de la Célula/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Proteínas Motoras Moleculares/metabolismo , Cadenas Pesadas de Miosina/metabolismo , Metástasis de la Neoplasia/tratamiento farmacológico , Saponinas/farmacología , Actinas/metabolismo , Animales , Línea Celular Tumoral , Femenino , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Ratones Desnudos , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
OBJECTIVE: To establish a method used for optimization of harvesting time and determine the best time for harvesting Rumex gmelini. METHOD: An HPLC method was applied to determinate the contents of seven active constituents(resveratrol, polydatin, chrysophanol 1-glucoside, nepodin, emodin, chrysophanol and physcion)of R. gmelini at different development stage. The result was analyzed by principal component analysis. RESULT: The accumulation of active constituents showed a regular pattern. CONCLUSION: The best harvesting time of R. gmelini is early July.
Asunto(s)
Emodina/análisis , Plantas Medicinales/química , Análisis de Componente Principal , Rumex/química , Antraquinonas/análisis , Emodina/análogos & derivados , Glucósidos/análisis , Raíces de Plantas/química , Plantas Medicinales/crecimiento & desarrollo , Análisis de Componente Principal/métodos , Control de Calidad , Resveratrol , Rumex/crecimiento & desarrollo , Estaciones del Año , Estilbenos/análisisRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Qianhu, the dried roots of Peucedanum praeruptorum DUNN (Umbelliferae), is a well-known traditional Chinese medicinal herb which was officially listed in the Chinese Pharmacopoeia. Praeruptorin D (PD) is one of the major active constituents of Peucedanum praeruptorum Dunn (Qianhu). The Pregnane X receptor (PXR) is an orphan nuclear receptor and plays a pivotal role in the activation of human cytochrome P450 3A4 (CYP3A4) gene. AIM OF THE STUDY: The purpose of this study was to investigate the effect of PD on the PXR-mediated transactivation of CYP3A4, and thus to predict potential herb-drug interactions between PD, Qianhu, and the other co-administered drugs that metabolized by CYP3A4. MATERIALS AND METHODS: The effect of PD on the Cyp3a11, mPXR mRNA expression in mice primary hepatocytes was measured using real-time PCR. The gene expression, protein expression, and catalytic activity of CYP3A4 in the LS174T cells after transfected with PXR expression plasmids were determined by real-time PCR, Western blot analysis, and LC-MS/MS based CYP3A4 substrate assay. RESULTS: The results revealed that the level of Cyp3a11 gene expression in mice primary hepatocytes was significantly increased by PD, but PD cannot induce the mPXR gene expression. On the other hand, CYP3A4 mRNA, protein expression and functional activity in PXR-over-expression LS174T cells were significantly increased by PD through PXR-mediated pathway; conversely, no significant change was found in the untransfected cells. CONCLUSIONS: These findings suggest that PD can significantly up-regulate CYP3A4 expression and activity via the PXR-mediated pathway and this should be taken into consideration to predict any potential herb-drug interactions when PD and Peucedanum praeruptorum Dunn are co-administered with other drugs.
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Apiaceae/química , Cumarinas/farmacología , Citocromo P-450 CYP3A/biosíntesis , Citocromo P-450 CYP3A/genética , Proteínas de la Membrana/genética , Extractos Vegetales/farmacología , Receptores de Esteroides/metabolismo , Animales , Línea Celular , Cumarinas/química , Citocromo P-450 CYP3A/metabolismo , Expresión Génica/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Interacciones de Hierba-Droga , Humanos , Masculino , Medicina Tradicional China/métodos , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/química , Receptor X de Pregnano , ARN Mensajero/genética , Receptores de Esteroides/genética , Activación Transcripcional , Regulación hacia Arriba/efectos de los fármacosRESUMEN
We report a case of severe hyponatremia related to duloxetine and ziprasidone. A 50-year-old woman on duloxetine and ziprasidone treatment for major depressive disorder developed polydipsia, polyuria, and two episodes of seizures, followed by admission to the emergency department on the 10th day of treatment. Laboratory investigations revealed elevated creatine kinase (CK) as well as hyponatremia, hypo-osmolality, and increased urine sodium. Syndrome of Inappropriate Antidiuretic Hormone (SIADH) was considered, although urine osmolality was not measured. Duloxetine and ziprasidone were discontinued and the CK gradually normalized after correction of hyponatremia. Clinicians should be aware of the possibility of antipsychotic-induced hyponatremia, particularly in patients with symptoms of polydipsia.
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Antidepresivos/efectos adversos , Antipsicóticos/efectos adversos , Trastorno Depresivo Mayor/tratamiento farmacológico , Hiponatremia/inducido químicamente , Piperazinas/efectos adversos , Tiazoles/efectos adversos , Tiofenos/efectos adversos , Creatina Quinasa/sangre , Clorhidrato de Duloxetina , Femenino , Humanos , Persona de Mediana EdadRESUMEN
BACKGROUND: Quercetin has been shown to induce apoptosis in a number of cancer cell lines, but a quercetin-loaded nanoliposomal formulation with enhanced antitumor activity in C6 glioma cells and its effect on cancer cell death has not been well studied. The aim of this study was to examine if quercetin-loaded liposomes (QUE-NL) has enhanced cytotoxic effects and if such effects involve type III programmed cell death in C6 glioma cells. METHODS: C6 glioma cells were treated with QUE-NL and assayed for cell survival, apoptosis, and necrosis. Levels of reactive oxygen species production and loss of mitochondrial membrane potential (ΔΨm) were also determined by flow cytometry assay to assess the effects of QUE-NL. ATP levels and lactate dehydrogenase activity were measured, and Western blotting was used to assay cytochrome C release and caspase expression. RESULTS: QUE-NL induced type III (necrotic) programmed cell death in C6 glioma cells in a dose-dependent and time-dependent manner. High concentrations of QUE-NL induced cell necrosis, which is distinct from apoptosis and autophagy, whereas liposomes administered alone induced neither significant apoptosis nor necrosis in C6 glioma cells. QUE-NL-induced ΔΨm loss and cytochrome C release had no effect on caspase activation, but decreased ATP levels and increased lactate dehydrogenase activity indicated that QUE-NL stimulated necrotic cell death. CONCLUSION: C6 glioma cells treated with QUE-NL showed a cellular pattern associated with necrosis without apoptosis and was independent of caspase activity. Nonapoptotic cell death induced by high concentrations of QUE-NL for controlling caspase-independent type III programmed cell death may provide the basis for novel therapeutic approaches to overcome avoidance of apoptosis by malignant cells.
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Glioma/tratamiento farmacológico , Liposomas/farmacología , Nanopartículas/química , Quercetina/farmacología , Adenosina Trifosfato/metabolismo , Animales , Antioxidantes/química , Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Línea Celular Tumoral , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocromos c/metabolismo , Glioma/patología , L-Lactato Deshidrogenasa/metabolismo , Liposomas/química , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Necrosis , Quercetina/química , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo C18 column (250 mm×4.6 mm, 5 µm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2%, v/v). Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 °C. An ultraviolet (UV) detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 µg/mL for 12, 13-dihydroxyeuparin (r>0.9999) and 106.9-1068.9 µg/mL for glycyrrhizic acid (r>0.9999), respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.
RESUMEN
AIMS: Investigation of the response of mesenchymal stem cells (MSCs) to vascular mechanical forces is very important in the field of cardiovascular intervention. Ser/Thr-protein kinase Pim-1 is a novel transducer of cell survival and the cell cycle that promotes signals in the hematopoietic cell system. Current studies aim to foster an understanding of Pim-1 expression and regulation in MSCs in response to different durations and strengths of laminar shear stress (SS) and to investigate the role of Pim-1 in SS-induced cell proliferation. MAIN METHODS: A parallel-plate flow chamber was used to control the strength and duration of SS. Proliferation was measured with the BrdU cell proliferation assay. The expressions of Pim-1 mRNA and protein were evaluated by reverse transcription-polymerase chain reaction and western blotting, respectively. RNA interference was used to knock down the Pim-1 gene. KEY FINDINGS: The results showed that SS up-regulation of Pim-1 mRNA and protein was time-dependent. Pim-1 induction was SS strength-dependent, and the expression level reached a maximum at 30 dynes/cm(2). Inhibitors of p38MAPK and ERK attenuated the SS-induced expression of Pim-1. In addition, SS significantly increased BrdU-uptake, which was effectively blocked by the silencing of Pim-1. SIGNIFICANCE: These results demonstrated that Pim-1 is expressed in MSCs and plays an important role in the SS-induced proliferation of MSCs.
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Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Proteínas Proto-Oncogénicas c-pim-1/metabolismo , Estrés Mecánico , Regulación hacia Arriba , Animales , Western Blotting , Proliferación Celular , Células Cultivadas , ARN Mensajero/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
A silica gel column chromatography method is established for the isolation and purification of euparin and 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis. For the first time, a high-performance liquid chromatography (HPLC) method is developed to determine simultaneously two benzofurans with UV absorptions at 240 nm. The analysis is performed on a Diamonsil C18 column with a gradient solvent system of acetonitrile and aqueous phosphoric acid (0.2%, v/v) at a flow-rate of 1.0 mL min⻹. All calibration curves reveal good linearity (r² > 0.9998) within the tested concentration ranges. The relative standard deviations for intra-day and inter-day are less than 2.0% and the recoveries range from 98.32% to 103.68% with relative SDs less than 2.0%. This method is successfully applied to quantitate two benzofurans in Radix Eupatorii Chinensis. Therefore, the new HPLC method is proven to be reliable and suitable for the quality control of Radix Eupatorii Chinensis.