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1.
Bioorg Chem ; 144: 107176, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38330721

RESUMEN

Repurposing drugs can significantly reduce the time and costs associated with drug discovery and development. However, many drug compounds possess intrinsic fluorescence, resulting in aberrations such as auto-fluorescence, scattering and quenching, in fluorescent high-throughput screening assays. To overcome these drawbacks, time-resolved technologies have received increasing attention. In this study, we have developed a rapid and efficient screening platform based on time-resolved emission spectroscopy in order to screen for inhibitors of the DNA repair enzyme, uracil-DNA glycosylase (UDG). From a database of 1456 FDA/EMA-approved drugs, sodium stibogluconate was discovered as a potent UDG inhibitor. This compound showed synergistic cytotoxicity against 5-fluorouracil-resistant cancer cells. This work provides a promising future for time-resolved technologies for high-throughput screening (HTS), allowing for the swift identification of bioactive compounds from previously overlooked scaffolds due to their inherent fluorescence properties.


Asunto(s)
Neoplasias de la Próstata , Uracil-ADN Glicosidasa , Humanos , Masculino , Uracil-ADN Glicosidasa/química , Oligonucleótidos , Gluconato de Sodio Antimonio , Evaluación Preclínica de Medicamentos , Reposicionamiento de Medicamentos , Detección Precoz del Cáncer
2.
Biomed Pharmacother ; 147: 112586, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34999373

RESUMEN

Extracellular vehicles have a natural targeting ability and immune tolerance of being usually applied in drug delivery systems; however, the purification of EVs is complicated and the production yield was quite low. We developed an artificial cellular mimetic nanovesicle (NV) with melanoma fragment membrane for the transportation with curcumin to achieve the anticancer purpose. B16F10 derived NVs were manufactured by the breakdown of cells using a series of extrusions through cut-off size filters (10 and 5 µm), and the whole procedure was easy and time-saving. To terminate the suspicion of cancer metastatic issue, B16F10 cells were treated by 30-min sonication and 1-min UVB exposure to remove genetic materials before the extrusion. B16F10 derived NV loaded with curcumin was called NV(S30U1/Cur), and the anticancer effect was evaluated by cell-based viability, immune, migration, and invasion. The results showed that NVs were manufactured by passing through 10 and 5 µm filters having an enviable production yield, and the mRNA amounts were declined within NVs produced by B16F10 cells treated with UVB in a comparison to the control group. NV(S30U1/Cur) were effectively decreased B1610 cell viability, and migratory and invasive abilities were also reduced significantly. Besides, CD8+ expression of murine primary lymphocytes was activated with CD4+ reduction by NV(S30U1/Cur) to stimulate the inherent tumor suppressive capacity in the immune system. Taken together, we established bioengineered NVs serving as novel cell mimetic nanocarriers to deliver natural compound for malignant melanoma potential immune chemotherapy. DATA AVAILABILITY STATEMENT: The data used to support the findings of this study are available from the corresponding author upon requests.


Asunto(s)
Antineoplásicos/farmacología , Curcumina/farmacología , Portadores de Fármacos , Melanoma/tratamiento farmacológico , Nanopartículas , Neoplasias Cutáneas/tratamiento farmacológico , Animales , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Línea Celular Tumoral/efectos de los fármacos , Curcumina/química , Curcumina/uso terapéutico , Sistemas de Liberación de Medicamentos , Ratones , Ratones Endogámicos C57BL
3.
Int J Biol Macromol ; 183: 1715-1722, 2021 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-34044030

RESUMEN

Radix Astragali polysaccharide RAP has been reported to play a crucial role in hematopoiesis without a clear mechanism. In this study, RAP's effects to enhance the recovery of cyclophosphamide (Cy)-suppressed bone marrow and blood cells is confirmed in vivo first. Confocal micrographs demonstrated the interesting direct binding of FITC-RAP to hematopoietic stem cells (HSC) in bone marrow. RAP protects both mice and human HSC in terms of cell morphology, proliferation, and apoptosis. RNA-sequencing and shRNA approaches revealed FOS to be a key regulator in RAP's protection. These evidences provide an unreported mechanism that RAP directly protects hematopoietic stem cells from chemotherapy-induced myelosuppression by increasing FOS expression.


Asunto(s)
Ciclofosfamida/efectos adversos , Medicamentos Herbarios Chinos/química , Hematopoyesis , Células Madre Hematopoyéticas/citología , Polisacáridos/administración & dosificación , Proteínas Proto-Oncogénicas c-fos/genética , Animales , Astragalus propinquus , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Femenino , Células Madre Hematopoyéticas/química , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Ratones , Polisacáridos/farmacología , Análisis de Secuencia de ARN , Regulación hacia Arriba
4.
J Pharm Biomed Anal ; 185: 113235, 2020 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-32182447

RESUMEN

Polysaccharides have broad bioactivities and are major components of water decoction of herb formulae. However, the quality control of polysaccharides remains a challenge. Oligosaccharide-fragment approach has been considered in elucidating chemical structures of polysaccharides, but never been used for quantitation. Using reference chemicals and a real sample Danggui Buxue Tang (DBT) in this study, an oligosaccharide-marker approach was established to quantify specific polysaccharides. Firstly, linear relationships between parent polysaccharides and hydrolysis-produced daughter oligosaccharides were verified using reference polysaccharides. Then in case of DBT, two fluorescence-labeled oligosaccharides with high specificity to individual parent polysaccharides were selected as markers. They were easily isolated and identified. Their potential in quantification of parent polysaccharides were satisfactorily validated in terms of linearity (r≥0.99), repeatability (RSD ≤ 8.4 %), and spike recovery (≥80 %). This method could be a promising approach for quality assessment of polysaccharides in herbal formulae.


Asunto(s)
Química Farmacéutica/métodos , Medicamentos Herbarios Chinos/análisis , Oligosacáridos/análisis , Control de Calidad , Química Farmacéutica/normas , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , Medicamentos Herbarios Chinos/química , Estándares de Referencia , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray/métodos , Espectrometría de Masa por Ionización de Electrospray/normas , Espectrometría de Masas en Tándem/métodos , Espectrometría de Masas en Tándem/normas
5.
Molecules ; 24(24)2019 Dec 04.
Artículo en Inglés | MEDLINE | ID: mdl-31817099

RESUMEN

As protein-protein interactions (PPIs) are highly involved in most cellular processes, the discovery of PPI inhibitors that mimic the structure of the natural protein partners is a promising strategy toward the discovery of PPI inhibitors. In this review, we discuss recent advances in the application of virtual screening for identifying mimics of protein partners. The classification and function of the mimicking protein partner inhibitor discovery by virtual screening are described. We anticipate that this review would be of interest to medicinal chemists and chemical biologists working in the field of protein-protein interaction inhibitors or probes.


Asunto(s)
Evaluación Preclínica de Medicamentos , Interfaz Usuario-Computador , Diseño de Fármacos , Descubrimiento de Drogas , Humanos , Unión Proteica , Mapeo de Interacción de Proteínas
6.
J Chromatogr A ; 1607: 460388, 2019 Dec 06.
Artículo en Inglés | MEDLINE | ID: mdl-31351593

RESUMEN

Qualitative and quantitative analysis of polysaccharides in herb formula remain challenge due to the limited choices of analytical methods concerning the intrinsic characteristics of large molecular mass. Herein, an oligosaccharide-marker approach was newly developed for quality assessment of polysaccharides in herbal materials, using Dendrobium officinale as a case study. This method involved partial acid hydrolysis of D. officinale polysaccharide (DOP) followed by p-aminobenzoic ethyl ester (ABEE) derivatization. Two ABEE-labeled oligosaccharides namely, Te-Man-ABEE and Pen-Man-ABEE, were selected as chemical markers due to their high specificity in herb formula. The linear relationship between the content of these two markers and the content of DOP was then successfully established respectively. The linear relationship was further transformed to that between peak area of chemical markers and DOP content so that chemical markers were not necessary to be isolated for analysis. This linear relationship was systemically validated in terms of precision and accuracy. The results showed that these two oligosaccharide-markers presented a good linear relationship with DOP (R2 ≥ 0.997) in the range of 0.68-16.02 µg. These markers also demonstrated satisfactory precision (RSD < 7.0%), and recovery (91.41%-118.30%) in real sample determination. Additionally, there was no significant difference between the results given by the two chemical markers as the RSD values were not more than 7.0%. While concerning the results given by the oligosaccharide-markers and the previously-published polysaccharide marker, the RSD value was not more than 6.4%. These suggest that the oligosaccharide-marker approach is a simple, quick, and reliable method to qualitatively and quantitatively determine of specific polysaccharide in herb formula.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dendrobium/química , Espectrometría de Masas/métodos , Oligosacáridos/química , Extractos Vegetales/química , Fluorescencia , Hidrólisis , Peso Molecular
7.
Anal Biochem ; 580: 49-55, 2019 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-31194944

RESUMEN

A G-quadruplex-based platform has been developed for the time-resolved monitoring of ochratoxin A (OTA). The simple platform displays good sensitivity for OTA with a detection limit of 40 nM via steady-state emission spectroscopy. Notably, the platform showed a detection limit of 10.8 nM via time-resolved emission spectroscopy (TRES), which is about 4 times more sensitive than steady-state mode. Moreover, the probe showed excellent selectivity for OTA over other mycotoxins. Furthermore, OTA was successfully detected in actual herbal plant extracts samples. Our platform is the first to detect OTA using TRES to distinguish between the target signals versus the auto-fluorescence of real samples. This platform shows improved detection speed, accuracy and sensitivity with simple operation, low cost, and no requirement for complicated pre-processing.


Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , G-Cuádruplex , Ocratoxinas/análisis , Iridio/química , Luminiscencia
8.
J Agric Food Chem ; 67(21): 5968-5977, 2019 May 29.
Artículo en Inglés | MEDLINE | ID: mdl-31037938

RESUMEN

Polysaccharides from functional foods have been proved to have diverse bioactivities, but little is known about what exactly happens to these polysaccharides after oral administration and even less about the underlying mechanism of action. Taking the marker polysaccharide (DOP) of Dendrobium officinale as an example, this study aims to demonstrate the dynamic distribution and degradation of orally dosed DOP in mice and in vitro using near-infrared fluorescence imaging and a kind of chromatographic analysis. The results indicate that (1) neither DOP nor fluorescence-labeled DOP (FDOP) was absorbed, (2) both DOP and FDOP were undigested and were quickly degraded to short-chain fatty acids in the large intestine, (3) DOP modulated gut microbiota, which could be associated with DOP's suppression of 4T1 tumor growth in mice. All of these findings suggest that some (maybe not all) bioactive polysaccharides share a common destiny: indigestible and nonabsorbing, ends in modulating bioactivities-associated gut microbiota.


Asunto(s)
Dendrobium/metabolismo , Microbioma Gastrointestinal , Extractos Vegetales/metabolismo , Polisacáridos/metabolismo , Animales , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Dendrobium/química , Femenino , Alimentos Funcionales/análisis , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/química , Polisacáridos/química
9.
Eur J Med Chem ; 143: 1021-1027, 2018 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-29232579

RESUMEN

NEDD8-activating enzyme (NAE) is an essential player of the NEDD8 conjugation pathway that regulates protein degradation. Meanwhile, drug repurposing is a cost-efficient strategy to identify new therapeutic uses for existing scaffolds. In this report, mitoxantrone (1) was repurposed as an inhibitor of NAE by virtual screening of an FDA-approved drug database. Compound 1 inhibited NAE activity in cell-free and cell-based systems with high selectivity and was competitive with ATP. Furthermore, compound 1 induced apoptosis of colorectal adenocarcinoma cancer cells through inhibiting the degradation of the neddylation substrate p53.


Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Neoplasias Colorrectales/tratamiento farmacológico , Inhibidores Enzimáticos/farmacología , Mitoxantrona/farmacología , Proteína NEDD8/antagonistas & inhibidores , Enzimas Activadoras de Ubiquitina/antagonistas & inhibidores , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Apoptosis/efectos de los fármacos , Células CACO-2 , Línea Celular , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Relación Dosis-Respuesta a Droga , Descubrimiento de Drogas , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Humanos , Mitoxantrona/síntesis química , Mitoxantrona/química , Modelos Moleculares , Estructura Molecular , Proteína NEDD8/metabolismo , Relación Estructura-Actividad , Proteína p53 Supresora de Tumor/antagonistas & inhibidores , Proteína p53 Supresora de Tumor/metabolismo , Enzimas Activadoras de Ubiquitina/metabolismo
10.
Molecules ; 22(1)2017 Jan 12.
Artículo en Inglés | MEDLINE | ID: mdl-28085113

RESUMEN

Sanhuang Tablet (SHT) is a Chinese patented drug commonly used for the treatment of inflammations of the respiratory tract, gastrointestinal tract, and skin. It contains a special medicinal composition including the single compound berberine hydrochloride, extracts of Scutellariae Radix and Rhei Radix et Rhizoma, as well as the powder of Rhei Radix et Rhizoma. Despite advances in analytical techniques, quantitative evaluation of a Chinese patented drug like SHT remains a challenge due to the complexity of its chemical profile. In this study, ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was used to simultaneously quantify 29 non-sugar small molecule components of SHT (11 flavonoids, two isoflavonoids, one flavanone, five anthraquinones, two dianthranones, five alkaloids, two organic acids and one stilbene). Three major saccharide components, namely fructose, glucose, and sucrose, were also quantitatively determined using high performance liquid chromatography-charged aerosol detector (HPLC-CAD) on an Asahipak NH2P-50 4E amino column. The established methods were validated in terms of linearity, sensitivity, precision, accuracy, and stability, and then successfully applied to analyze 27 batches of commercial SHT products. A total of up to 57.61% (w/w) of SHT could be quantified, in which the contents of the determined non-saccharide small molecules varied from 5.91% to 16.83% (w/w) and three saccharides accounted for 4.41% to 48.05% (w/w). The results showed that the quality of the commercial products was inconsistent, and only four of those met Chinese Pharmacopoeia criteria.


Asunto(s)
Medicamentos Herbarios Chinos/química , Inflamación/tratamiento farmacológico , Comprimidos/química , Alcaloides/química , Alcaloides/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/análisis , Flavonoides/química , Flavonoides/aislamiento & purificación , Humanos , Inflamación/patología , Medicina Tradicional de Asia Oriental , Rizoma/química , Scutellaria baicalensis/química , Comprimidos/análisis , Espectrometría de Masas en Tándem
11.
Molecules ; 22(1)2017 Jan 18.
Artículo en Inglés | MEDLINE | ID: mdl-28106808

RESUMEN

Dendrobium officinale extract shows potent anti-fatigue effects; however, the active substance responsible for these effects remains undetermined. A glucomannan with a huge molecular size of 730 kDa, called DOP, was identified as the unique authentication marker of this expensive herb. DOP exhibited immunomodulating effects on macrophages and lymphocytes in our previous study. Clinical reports also showed that people with fatigue syndrome have a disturbed immune system. Because DOP is the unique and dominant component of D. officinale, we hypothesize that DOP may also have anti-fatigue activity. The present study aims to evaluate the anti-fatigue activity of DOP on BALB/c mice, with Rhodiola rosea extract as a positive control. DOP and Rhodiola rosea extract were orally administered at doses of 50 mg/kg and 100 mg/kg, respectively, for four weeks, and the anti-fatigue activity of DOP on BALB/c mice was evaluated using the weight-loaded swimming test. The contents of lactic dehydrogenase (LDH), creatine phosphokinase (CK), triglyceride (TG), blood urea nitrogen (BUN), superoxide dismutase (SOD), malondialdehyde (MDA), lactic acid (LD), and glutathione peroxidase (GSH-Px) in serum, glycogen of liver and gastrocnemius muscle were also determined. Their effects on variability of T cells and B cells were determined by using tetrazolium compound (MTS) method. The weight-loaded swimming exercise caused fatigue syndrome, mainly including the decreases of serum SOD/GSH-Px and gastrocnemius glycogen, as well as the increases of LDH, BUN, MDA, CK, TG, and LD in serum. All of these indicators of fatigue were inhibited to a certain extent by both DOP and Rhodiola rosea extract; however, the effects of DOP were much stronger than those of Rhodiola rosea extract. Compared to the positive control, mice dosed with DOP showed increases in endurance, body weight, and food intake. Furthermore, DOP-feeding mice significantly increased the cell variability of T lymphocytes and B lymphocytes, compared with that of mice in control group. This study indicates that the unique and dominant polysaccharide DOP of D. officinale has stronger anti-fatigue activity than Rhodiola rosea extract. As such, DOP has promising potential for pharmaceutical development into health products to reduce fatigue.


Asunto(s)
Anabolizantes/farmacología , Dendrobium/química , Ingestión de Alimentos/efectos de los fármacos , Fatiga/prevención & control , Resistencia Física/efectos de los fármacos , Polisacáridos/farmacología , Anabolizantes/aislamiento & purificación , Animales , Linfocitos B/citología , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Nitrógeno de la Urea Sanguínea , Peso Corporal/efectos de los fármacos , Creatina Quinasa/sangre , Ingestión de Alimentos/fisiología , Fatiga/sangre , Fatiga/fisiopatología , Glutatión Peroxidasa/sangre , L-Lactato Deshidrogenasa/sangre , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Malondialdehído/antagonistas & inhibidores , Malondialdehído/metabolismo , Ratones , Ratones Endogámicos BALB C , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Condicionamiento Físico Animal , Extractos Vegetales/química , Polisacáridos/aislamiento & purificación , Rhodiola/química , Superóxido Dismutasa/sangre , Natación , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Triglicéridos/sangre
12.
Am J Chin Med ; 45(1): 123-136, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28081631

RESUMEN

Baicalein (BA), one of the major compounds isolated from the root of Scutellaria baicalensis Gerogi, exhibits various pharmacological effects, such as anti-oxidant, anti-inflammatory, and anticancer effects. In this study, we found that BA reduced cell viability and increased apoptosis in ovarian cancer cells. Treatment of cells with BA enhanced microtubule-associated protein light chain 3-II (LC3-II) expression, acidic vesicular organelle and GFP-LC3 fluorescence dot accumulation. Combined treatment with chloroquine and BA apparently reduced cell viability and increased the cleavage of poly (ADPribose) polymerase (PARP) in both HEY and A2780 ovarian cancer cell lines, indicating that BA induces a protective autophagy in these cells. Knockdown of Beclin 1 by siRNA remarkably decreased BA-induced LC3-II lipidation. In addition, we found an increase in the phosphorylation of extracellular signal-regulated kinase (ERK, Thr202/Thr204) and AKT (Ser473) after BA treatment, and inhibition of ERK activation by the pharmacological inhibitor U0126 or ERK siRNA blocked BA-induced autophagy. Taken together, these results suggest that BA induces Beclin 1- and ERK-dependent autophagy in ovarian cancer cells.


Asunto(s)
Autofagia/efectos de los fármacos , Beclina-1/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Flavanonas/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas Asociadas a Microtúbulos/efectos de los fármacos , Antimaláricos/farmacología , Beclina-1/metabolismo , Western Blotting , Línea Celular Tumoral , Cloroquina/farmacología , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Técnicas In Vitro , Microscopía Confocal , Microscopía Fluorescente , Proteínas Asociadas a Microtúbulos/metabolismo , Neoplasias Ováricas , Poli(ADP-Ribosa) Polimerasas/efectos de los fármacos , Poli(ADP-Ribosa) Polimerasas/metabolismo , ARN Interferente Pequeño
13.
Phytomedicine ; 23(9): 939-47, 2016 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-27387402

RESUMEN

BACKGROUND: Psoralidin (PSO), a natural phenolic coumarin, was reported to have anti-cancer activities. PSO induced reactive oxygen species (ROS) generation in cancer cells. The role of ROS in its anti-cancer effect remains unclear. PURPOSE: This study was designed to investigate the potential roles of ROS in PSO-induced anti-cancer effect in MCF-7 breast cancer cells. METHODS: Effect of PSO on cancer cell proliferation was determined by MTT assay. Comet assay was used to determine DNA damage. Protein expression was detected by Western blotting. Autophagic vacuoles were detected by monodansylcadaverine (MDC) staining. ROS generation was measured by fluorescent probe. NOX4 localization was determined by immunofluorescence staining. RESULTS: PSO treatment caused proliferation inhibition in time- and dose- dependent manners, which was partially reversed by N-acetyl cysteine (NAC) and diphenyleneiodonium (DPI). PSO induced DNA damage and increased protein expression of γ-H2AX, phosphorylation of ATM, ATR, Chk1, and Chk2. PSO induced autophagy as evidenced by the accumulation of autophagic vacuoles and alterations of autophagic protein expression. PSO-induced cell death was enhanced by autophagy inhibitor chloroquine (CQ). Furthermore, PSO treatment induced ROS formation, which was reversed by NAC or DPI pretreatment. The expression of NOX4 was significantly enhanced by PSO. Both NAC and DPI could reverse PSO-induced DNA damage and autophagic responses. In addition, silencing NOX4 by siRNA inhibited PSO-induced ROS generation, DNA damage, and autophagy. CONCLUSIONS: Taken together, these results showed that PSO induced DNA damage and protective autophagy mediated by ROS generation in a NOX4-dependent manner in MCF-7 cells.


Asunto(s)
Antineoplásicos Fitogénicos/toxicidad , Autofagia/efectos de los fármacos , Benzofuranos/toxicidad , Neoplasias de la Mama/genética , Cumarinas/toxicidad , Daño del ADN , NADPH Oxidasas/metabolismo , Especies de Nitrógeno Reactivo , Apoptosis/efectos de los fármacos , Cadaverina/análogos & derivados , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Colorantes Fluorescentes , Silenciador del Gen , Humanos , Células MCF-7 , NADPH Oxidasa 4 , NADPH Oxidasas/genética , ARN Interferente Pequeño/farmacología
14.
Oxid Med Cell Longev ; 2016: 2853543, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27403230

RESUMEN

Equisetum ramosissimum, a genus of Equisetaceae, is a medicinal plant that can be separated into ethyl acetate (EA), dichloromethane (DM), n-hexane (Hex), methanol (MeOH), and water extracts. EA extract was known to have potent antioxidative properties, reducing power, DPPH scavenging activity, and metal ion chelating activity. This study compared these five extracts in terms of their inhibiting effects on three human malignant melanomas: A375, A375.S2, and A2058. MTT assay presented the notion that both EA and DM extracts inhibited melanoma growth but did not affect the viabilities of normal dermal keratinocytes (HaCaT) or fibroblasts. Western blot analyses showed that both EA and DM extracts induced overexpression of caspase proteins in all three melanomas. To determine their roles in melanogenesis, this study analyzed their in vitro suppressive effects on mushroom tyrosinase. All extracts except for water revealed moderate suppressive effects. None of the extracts affected B16-F10 cells proliferation. EA extract inhibited cellular melanin production whereas DM extract unexpectedly enhanced cellular pigmentation in B16-F10 cells. Data for modulations of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein 1, and tyrosinase-related protein 2 showed that EA extract inhibited protein expression mentioned above whereas DM extract had the opposite effect. Overall, the experiments indicated that the biofunctional activities of EA extract contained in food and cosmetics protect against oxidation, melanoma, and melanin production.


Asunto(s)
Equisetum/química , Melanocitos/efectos de los fármacos , Extractos Vegetales/química , Animales , Humanos , Melanoma Experimental , Ratas , Especies Reactivas de Oxígeno
15.
J Pharm Biomed Anal ; 125: 415-26, 2016 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-27131804

RESUMEN

YinHuang drop pill (YHDP) is a new preparation, derived from the traditional YinHuang (YH) decoction. Since drop pills are one of the newly developed forms of Chinese patent drugs, not much research has been done regarding the quality and efficacy. This study aims to establish a comprehensive quantitative analysis of the chemical profile of YHDP. ultra high-performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS/MS) was used to identify 34 non-sugar small molecules including 15 flavonoids, 9 phenolic acids, 5 saponins, 1 iridoid, and 4 iridoid glycosides in YHDP samples, and 26 of them were quantitatively determined. Sugar composition of YHDP in terms of fructose, glucose and sucrose was examined via a high performance liquid chromatography-evaporative light scattering detector on an amide column (HPLC-NH2P-ELSD). Macromolecules were examined by high performance gel permeation chromatography coupled with ELSD (HPGPC-ELSD). The content of the drop pill's skeleton component PEG-4000 was also quantified via ultra-high performance liquid chromatography coupled with charged aerosol detector (UHPLC-CAD). The results showed that up to 73% (w/w) of YHDP could be quantitatively determined. Small molecules accounted for approximately 5%, PEG-4000 represented 68%, while no sugars or macromolecules were found. Furthermore, YHDP showed no significant differences in terms of daily dosage, compared to YinHuang granules and YinHuang oral liquid; however, it has a higher small molecules content compared to YinHuang lozenge.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos , Espectrometría de Masas/métodos
16.
J Ethnopharmacol ; 179: 243-52, 2016 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-26743224

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Polysaccharides of Radix Astragali (Astragalus membranaceus (Fisch) Bge.; Huangqi) are able to induce cytokine production of macrophages and are considered the main active ingredient for the immune-enhancing effect of this commonly used medicinal herb. AIM OF STUDY: To investigate the molecular mechanism of immunomodulating activities of a reported Astragalus polysaccharide, RAP, which is a hyperbranched heteroglycan with average molecular weight of 1334kDa. MATERIALS AND METHODS: The cytokine production of RAW264.7 cells were analyzed by using ELISA assays while cell viability was assessed by MTT method. Western blot analysis was used for determining protein contents of mitogen-activated protein kinases (MAPKs). In addition, the level of IL-6, iNOS, and TNF-α mRNA was determined by RT-PCR. RESULTS: It has been found that RAP itself did not have any cytotoxic effect on mouse mammary carcinoma 4T1 cells, but it significantly enhanced cytotoxicity of the supernatant of RAW264.7cells on 4T1 cells. Furthermore, RAP enhanced the production of NO and cytokines in RAW264.7 cells, and significantly up-regulated gene expressions of TNF-α, IL-6, iNOS. All these bioactivities were blocked by the inhibitor of TLR4 (Toll-like receptor 4), suggesting that TLR4 is a receptor of RAP and mediates its immunomodulating activity. Further analyses demonstrated that RAP rapidly activated TLR4-related MAPKs, including phosphorylated ERK, phosphorylated JNK, and phosphorylated p38, and induced translocation of NF-κB as well as degradation of IκB-α. These results are helpful to better understand the immunomodulating effects of Radix Astragali. CONCLUSIONS: RAP may induce cytokine production of RAW264.7 cells through TLR4-mediated activation of MAPKs and NF-κB.


Asunto(s)
Planta del Astrágalo , Citocinas/metabolismo , Polisacáridos/farmacología , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismo , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Óxido Nítrico/metabolismo , Fosforilación/efectos de los fármacos , Polisacáridos/aislamiento & purificación , Receptor Toll-Like 4/antagonistas & inhibidores
17.
J Agric Food Chem ; 64(4): 881-9, 2016 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-26752248

RESUMEN

A crude polysaccharide fraction (cDOP) has been determined to be the characteristic marker of Dendrobium officinale, an expensive tea material in Asia, but its chemistry and bioactivity have not been studied. In work reported here, cDOP was destarched (DOP, 90% yield) and separated into two subfraction polysaccharides, DOPa and DOPb, which were characterized by monosaccharide composition and methylation analyses and spectral analyses (FT-IR and (1)H and (13)C NMR). Both are composed of mannose and glucose at similar ratios and have a similar structure with a backbone of 1,4-linked ß-D-mannopyranosyl and ß-D-glucopyranosyl residues. Significant differences were observed only in their molecular weights. Bioassay using mouse macrophage cell line RAW264.7 indicated that DOP and its two subfractions enhance cell proliferation, TNF-α secretion, and phagocytosis in a dose-dependent manner. They also induced the proliferation of lymphocytes alone and with mitogens. DOPa and DOPb are thus proven to be major, active polysaccharide markers of D. officinale.


Asunto(s)
Dendrobium/química , Factores Inmunológicos/química , Factores Inmunológicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Polisacáridos/química , Polisacáridos/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Factores Inmunológicos/aislamiento & purificación , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Fagocitosis/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Polisacáridos/aislamiento & purificación , Células RAW 264.7 , Factor de Necrosis Tumoral alfa/inmunología
18.
Am J Chin Med ; 43(6): 1265-79, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26394653

RESUMEN

Tanshinones are a group of bioactive constituents isolated from Salvia miltiorrhiza Bunge, a widely prescribed traditional Chinese herb. In the current study, the anticancer properties of total tanshinones (TDT) were evaluated using 95D lung cancer cells. Tanshinone IIA was identified as the main component of TDT. Compared with tanshinone IIA, TDT showed more cytotoxic effects on the 95D cells. Annexin V/7-AAD double staining, the depolarization of mitochondrial membrane potential (MMP) (Δψ), the up-regulation of pro-apoptotic proteins, such as cleaved-PARP, cleaved-caspase-3, Bax, and Bad, and the down-regulation of anti-apoptotic protein Bcl-2 were evidence of TDT-induced apoptosis. Furthermore, TDT-induced autophagy as demonstrated by monodansylcadaverine (MDC) staining and the up-regulation of autophagy-associated proteins, such as LC3-II, Beclin-1, Atg3, Atg5, Atg7, and Atg12. Autophagy inhibitors, 3-methyladenine (3-MA) and bafilomycin A1, enhanced TDT-induced cell death. 3-MA pretreatment enhanced the TDT-induced up-regulation of Bax and cleaved-PARP. In addition, TDT induced the generation of reactive oxygen species (ROS), which was reversed by N-acetylcysteine (NAC). NAC also reversed TDT-induced depolarization of Δψ, MDC staining, up-regulation of Bax, cleaved-PARP, Beclin-1, LC3-II, and cell viability. In conclusion, our findings showed that TDT-induced apoptosis and protective autophagy in 95D cells mediated by increasing intracellular ROS production.


Asunto(s)
Abietanos/farmacología , Autofagia/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Neoplasias Pulmonares/fisiopatología , Especies Reactivas de Oxígeno/metabolismo , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Beclina-1 , Caspasa 3/genética , Caspasa 3/metabolismo , Línea Celular Tumoral , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Regulación hacia Arriba
19.
ScientificWorldJournal ; 2015: 303415, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25839053

RESUMEN

Cinnamomum osmophloeum Kanehira belongs to the Lauraceae family of Taiwan's endemic plants. In this study, C. osmophloeum Kanehira extract has shown inhibition of tyrosinase activity on B16-F10 cellular system first. Whether extracts inhibited mushroom tyrosinase activity was tested, and a considerable inhibition of mushroom tyrosinase activity by in vitro assays was presented. Animal experiments of C. osmophloeum Kanehira were carried out by observing animal wound repair, and the extracts had greater wound healing power than the vehicle control group (petroleum jelly with 8% DMSO, w/v). In addition, the antioxidant capacity of C. osmophloeum Kanehira extracts in vitro was evaluated. We measured C. osmophloeum Kanehira extract's free radical scavenging capability, metal chelating, and reduction power, such as biochemical activity analysis. The results showed that a high concentration of C. osmophloeum Kanehira extract had a significant scavenging capability of free radical, a minor effect of chelating ability, and moderate reducing power. Further exploration of the possible physiological mechanisms and the ingredient components of skincare product for skin-whitening, wound repair, or antioxidative agents are to be done.


Asunto(s)
Antioxidantes/farmacología , Cinnamomum , Monofenol Monooxigenasa/antagonistas & inhibidores , Extractos Vegetales/farmacología , Cicatrización de Heridas/efectos de los fármacos , Animales , Antioxidantes/aislamiento & purificación , Evaluación Preclínica de Medicamentos/métodos , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/farmacología , Masculino , Melanoma Experimental , Monofenol Monooxigenasa/metabolismo , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta , Ratas , Ratas Wistar , Cicatrización de Heridas/fisiología
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