RESUMO
Estrogens and their analogues can cause harm to human health through the food chain. Ten estrogens in different milk samples were directly extracted by amphiphilic divinylbenzene/N-vinyl-2-pyrrolidone (DVB/NVP)-Fe3O4@SiO2-based magnetic solid-phase extraction (MSPE) followed by pre-column derivatization and ultra-high performance liquid chromatography tandem mass-spectrometry (UHPLC-MS/MS) detection. Under the optimal conditions, the limits of detection for ten analytes were in the range of 0.05-0.38 ng mL-1 in whole liquid milk matrix and 0.04-3.00 ng g-1 in milk powder matrix. The intra-/inter-day accuracy ranged in 83.4-113.8%, with RSDs in 2.5-15.0%. A total of 15 brands of liquid milk and milk powder samples were analyzed, and only estradiol was detected in three brands of boxed liquid milk within safe range. The proposed sample pretreatment eliminated the common protein precipitation process, improved the sample throughput, and has the potential for routine testing of estrogens and their analogues in market-sale milk samples.
RESUMO
The dyeing and adulteration of traditional Chinese medicines (TCMs) are continuously updated. Valuable analytical methods for the daily inspection of illegal colorant additives in TCMs and the preparations are in demand. Two deep eutectic solvent (DES)-based vortex-assisted liquid-liquid microextraction (VA-LLME) and ultrasonic-assisted solid-liquid microextraction (UA-SLME) were developed for the sample pretreatment of ten water-soluble colorants and five water-insoluble colorants, respectively, followed by an HPLC-DAD detection. Fifteen colorants were analyzed at four detection wavelengths within 40 min of gradient elution. The optimal DES of VA-LLME and UA-SLME were screened from 23 homemade DESs. The factors affecting the extraction efficiency of VA-LLME and UA-SLME were optimized systematically. Under the optimal conditions, ten water-soluble colorants analyzed by DES-based VA-LLME-HPLC-DAD showed good linearity (R ≥ 0.9995) within the optimal linear range. The LODs and LOQs were 0.2-1.0 µg g-1 and 0. 5-5.0 µg g-1, respectively. The recoveries of spiked samples were 80.2%-104.7 %, with RSDs ≤ 4.39 %. Five water-insoluble colorants of Sudan IâIV and Sudan 7B analyzed by DES-based UA-SLME-HPLC-DAD showed good linearity (R ≥ 0.9995) within the optimal linear range. The LODs and LOQs were 0.8-8.0 µg g-1 and 4.0-40.0 µg g-1, respectively. The recoveries of spiked samples were 94.2%-103.1 %, with RSDs ≤ 4.81 %. The proposed DES-based VA-LLME-HPLC-DAD was successfully applied to analyze six water-soluble yellow colorants in Cuscutae Semen, salted Cuscutae Semen, and four water-soluble red colorants in Schisandrae Chinensis Fructus. The proposed DES-based UA-SLME-HPLC-DAD was successfully applied to analyze five water-insoluble red colorants in Dieda pills. The study provides analytical method options for routine tests of water-soluble, water-insoluble, or both water-soluble/-insoluble illegal colorant additives in herbal medical materials and preparations by the relevant proposed DES-based sample pretreatment method or a combination of the two proposed DES-based methods.
Assuntos
Corantes , Medicamentos de Ervas Chinesas , Interações Hidrofóbicas e Hidrofílicas , Microextração em Fase Líquida , Cromatografia Líquida de Alta Pressão/métodos , Microextração em Fase Líquida/métodos , Corantes/química , Corantes/análise , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/análise , Medicina Tradicional Chinesa , Solventes/químicaRESUMO
Steroids are tetracyclic aliphatic compounds, and most of them contain carbonyl groups. The disordered homeostasis of steroids is closely related to the occurrence and progression of various diseases. Due to high structural similarity, low concentrations in vivo, poor ionization efficiency, and interference from endogenous substances, it is very challenging to comprehensively and unambiguously identify endogenous steroids in biological matrix. Herein, an integrated strategy was developed for the characterization of endogenous steroids in serum based on chemical derivatization, ultra-performance liquid chromatography quadrupole Exactive mass spectrometry (UPLC-Q-Exactive-MS/MS), hydrogen/deuterium (H/D) exchange, and a quantitative structure-retention relationship (QSRR) model. To enhance the mass spectrometry (MS) response of carbonyl steroids, the ketonic carbonyl group was derivatized by Girard T (GT). Firstly, the fragmentation rules of derivatized carbonyl steroid standards by GT were summarized. Then, carbonyl steroids in serum were derivatized by GT and identified based on the fragmentation rules or by comparing retention time and MS/MS spectra with those of standards. H/D exchange MS was utilized to distinguish derivatized steroid isomers for the first time. Finally, a QSRR model was constructed to predict the retention time of the unknown steroid derivatives. With this strategy, 93 carbonyl steroids were identified from human serum, and 30 of them were determined to be dicarbonyl steroids by the charge number of characteristic ions and the number of exchangeable hrdrogen or comparing with standards. The QSRR model built by the machine learning algorithms has an excellent regression correlation, thus the accurate structures of 14 carbonyl steroids were determined, among which three steroids were reported for the first time in human serum. This study provides a new analytical method for the comprehensive and reliable identification of carbonyl steroids in biological matrix.
Assuntos
Esteroides , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas em Tandem/métodos , Deutério , Cromatografia Líquida de Alta Pressão/métodos , Esteroides/análise , Cromatografia LíquidaRESUMO
Pharmaceuticals are widespread in aquatic ecosystem, which may pose a potential threat to fish and human. In the study, a robust and reliable magnetic solid-phase extraction (MSPE) based on bovine serum albumin (BSA) restricted access octadecyl/phenyl-mixed-functionalized magnetic silica nanoparticles (BSA-C18/Ph-Fe3O4@SiO2 NPs) as sorbent was developed for the extraction of venlafaxine, paroxetine, fluoxetine, norfluoxetine, sertraline and diphenhydramine from the muscle extracts of aquatic products followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection. The homemade sorbent showed appropriate compatibility in aqueous solution and good performance in reducing matrix interference of snakehead muscle tissue extracts with absolute matrix effect ranging in 95.4%-105.5%. The protocol was validated in analyte-free snakehead muscle with favorable recoveries ranging in 91.6%-103.6% and relative standard deviations (RSDs) less than 9.5%. Limits of detection (LODs) of the drugs were lower than 0.018 µg kg-1. Short-term drug-exposure experiments at low and high doses were conducted on snakeheads, and the measured contents of analytes were in the range 0.029-9.58 µg kg-1 with appropriate recoveries (90.0%-114.0%). The approach was extensively applied for the analysis of twelve species of market-sale aquatic products (total 37 samples), and up to 1.868 and 0.521 µg kg-1 of diphenhydramine and venlafaxine were measured, respectively. The approach shows remarkable potential in biological complex samples.
Assuntos
Antidepressivos/análise , Contaminação de Alimentos/análise , Nanopartículas de Magnetita , Alimentos Marinhos/análise , Dióxido de Silício , Animais , Cromatografia Líquida de Alta Pressão , Ecossistema , Limite de Detecção , Fenômenos Magnéticos , Soroalbumina Bovina , Extração em Fase Sólida , Espectrometria de Massas em TandemRESUMO
In this study, conventional Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method was modified by magnetic solid-phase extraction (MSPE) for purification/pre-concentration of eleven estrogens and estrogen mimics from the extracts of pork and chicken muscles, prior to dansyl chloride (DNS-Cl) derivatization coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) assay. Dual octadecyl- and 2-aminoethyl-3-aminopropyl- groups functionalized mesoporous silica core-shell magnetic nanoparticles (C18/NH2-Fe3O4@mSiO2 MNPs) were synthesized and employed as MSPE sorbent with remarkable aqueous compatibility in comparison with conventional C18 functionalized sorbent. The proposed MSPE is easier to handle than the traditional SPE purification process in QuEChERS method. The lab-prepared MNPs were characterized by transmission electron microscope (TEM), brunner-emmet-teller (BET), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), thermo-gravimetric analysis (TGA) and vibrating sample magnetometer (VSM). Pre-column derivatization was conducted to significantly enhance the sensitivity of the analytes in MS/MS via analyzing their derivatives in positive ion mode instead of analyzing their original forms in negative ion mode. Under the optimal sample pretreatment and instrumental analysis conditions, the approach showed low limits of detection (LODs, 0.02â3.00⯵g kg-1), appropriate recoveries (81.1â115.4%) and acceptable precisions (0.48â15.1%, nâ¯=â¯6), with good feasibility and future prospect of trace compounds analysis in complex food samples.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Estrogênios/análise , Magnetismo , Carne Vermelha/análise , Extração em Fase Sólida/economia , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos , Adsorção , Animais , Galinhas , Concentração de Íons de Hidrogênio , Limite de Detecção , Nanopartículas de Magnetita/ultraestrutura , Concentração Osmolar , Dióxido de Silício/química , Solventes , Espectroscopia de Infravermelho com Transformada de Fourier , Suínos , Fatores de TempoRESUMO
In the study, novel polysulfone hollow fiber membrane (PSF) supported polar enhanced phase (PEP) particles-polyacrylonitrile (PAN) polymer thin-film rods (PEP-PAN@PSF rods) were developed for the extraction of five steroidal endocrine disrupting chemicals (EDCs) (estrone (E1), estradiol (E2), estriol (E3), ethinyloestradiol (EE2), 2-methoxyestradiol (2ME2)) and four phenolic EDCs (bisphenol A (BPA), hexestrol (HEX), diethylstilbestrol (DES), dienestrol (DE)) in environmental water, sediment, and fish muscle homogenates, followed by pre-column derivatization and ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The traditional preparation method for thin-film rods was improved by introducing a supporting membrane PSF between the external thin-film coating and the internal bare rod, avoiding the conventional pre-corrosion by strong acid/base. The commercial PEP prepared rods showed competitive capacity for both polar and nonpolar EDCs. In addition, pre-column derivatization with dansyl chloride (DNS-Cl) was adopted for the phenolic analytes prior to UHPLC-MS/MS detection, leading to a significant enhancement of sensitivity via analyzing the dansylated derivatives under positive electrospray ionization (ESI) mode instead of the analytes under negative ESI mode. The protocol was validated in four matrices including environmental water, sediment and two fish species. No matrix effects were observed in four matrices. The limits of detection (LODs) for the analytes were in the range of 0.002-0.072 µg L-1 for environmental water, 0.032-0.734 ng g-1 for sediment, and 0.011-0.435 ng g-1 for two fish species, respectively. Appropriate linearity was observed for all the analytes with correlation coefficients (R2) above 0.997. The intra-day trueness of the approach at low, medium and high levels was in the range of 86.6-116.1% with relative standard deviations (RSDs) lower than 15.4%. And the inter-day trueness was in the range of 84.6-114.8% with RSDs lower than 16.3%. The proposed method was successfully applied for the analysis of nine EDCs in environmental water, sediment, and fish muscle homogenates.
Assuntos
Resinas Acrílicas/química , Monitoramento Ambiental , Sedimentos Geológicos/análise , Polímeros/química , Sulfonas/química , Poluentes Químicos da Água/análise , 2-Metoxiestradiol/análise , Animais , Cromatografia Líquida de Alta Pressão , Estradiol/análise , Estriol/análise , Estrona/análise , Etinilestradiol/análise , Peixes , Espectrometria de Massas em TandemRESUMO
Aim: Routine therapeutic drug monitoring is highly recommended since common antidepressant combinations increase the risk of drug-drug interactions or overlapping toxicity. Materials & methods: A magnetic solid-phase extraction by using C18-functionalized magnetic silica nanoparticles (C18-Fe3O4@SiO2 NPs) as sorbent was proposed for rapid extraction of venlafaxine, paroxetine, fluoxetine, norfluoxetine and sertraline from clinical plasma and urine samples followed by ultra-HPLC-MS/MS assay. Results: The synthesized C18-Fe3O4@SiO2 NPs showed high magnetization and efficient extraction for the analytes. After cleanup by magnetic solid-phase extraction, no matrix effects were found in plasma and urine matrices. The analytes showed LODs among 0.15-0.75 ng ml-1, appropriate linearity (R ≥ 0.9990) from 2.5 to 1000 ng ml-1, acceptable accuracies 89.1-110.9% with precisions ≤11.0%. The protocol was successfully applied for the analysis of patients' plasma and urine samples. Conclusion: It shows high potential in routine therapeutic drug monitoring of clinical biological samples.
Assuntos
Antidepressivos/análise , Nanopartículas de Magnetita/química , Antidepressivos/isolamento & purificação , Antidepressivos/metabolismo , Cromatografia Líquida de Alta Pressão , Fluoxetina/análogos & derivados , Fluoxetina/sangue , Fluoxetina/isolamento & purificação , Fluoxetina/urina , Humanos , Concentração de Íons de Hidrogênio , Limite de Detecção , Paroxetina/sangue , Paroxetina/isolamento & purificação , Paroxetina/urina , Reprodutibilidade dos Testes , Sertralina/sangue , Sertralina/isolamento & purificação , Sertralina/urina , Dióxido de Silício/química , Extração em Fase Sólida , Solventes/química , Espectrometria de Massas em Tandem , Cloridrato de Venlafaxina/sangue , Cloridrato de Venlafaxina/isolamento & purificação , Cloridrato de Venlafaxina/urinaRESUMO
A three-step strategy was developed for systematic characterization of curcuminoids in turmeric. Based on UHPLC-QTOF-MS/MS analysis, 89 curcuminoids including 16 novel ones were identified in the turmeric samples using this approach. During the identification process, false positive results were excluded by combining the positive and negative ESI-MS/MS analyses. Moreover, the characterization of the keto and enol forms of type A, B and C curcuminoids was first discussed and they were clearly distinguished using negative ESI-MS/MS method with UV spectra analyses. The structures of detected curcuminoids were identified and rationalized in both ion modes. Additionally, the fragmentation behaviors of the 15 types of curcuminoids were clearly illustrated in this work, which will be helpful for detection and identification of corresponding trace curcuminoids in complex turmeric samples using UHPLC-QTOF-MS/MS methods.
Assuntos
Técnicas de Química Analítica/métodos , Cromatografia Líquida de Alta Pressão , Curcuma/química , Espectrometria de Massas em TandemRESUMO
Turmeric and curcuminoids are used as natural food coloring and functional food additives in various parts of the world. In this study, ninety-six curcuminoids were fully characterized using a targeted curcuminoid profile, which established by integrated use of two complementary LC-MS/MS platforms (liquid chromatography-quadrupole time of flight mass spectrometry (LC-QTOF-MS/MS) and liquid chromatography-quadrupole linear ion trap mass spectrometry (LC-QTRAP-MS/MS)). The curcuminoid profile was represented in the form of a multiple reaction monitoring (MRM) mode based on LC-QTRAP-MS/MS analysis. It facilitated the qualitative and relative quantitative analysis of curcuminoids in a single injection. Meanwhile, the profile was successfully applied to the quality evaluation of raw materials of turmeric from different regions in China and Myanmar. The structural identification procedures of curcuminoids and the integrated strategy provide a suitable method to analyze targeted plant metabolites which occur in a high number but sharing either structural similarities or similar functional groups.
Assuntos
Cromatografia Líquida/métodos , Curcuma/química , Espectrometria de Massas em Tandem/métodos , China , Análise de Alimentos/métodos , Mianmar , Reprodutibilidade dos TestesRESUMO
This study investigated the effect of diosgenin, a natural sapogenin possessing various pharmacological activities, on benign prostatic hyperplasia (BPH) in rats and the possible mechanisms. BPH was established in the castrated rats by subcutaneous injection of testosterone propionate. Animals were randomly divided into four groups (n=10 each): model group (0.5% sodium carboxymethyl cellulose); positive control group (3 mg/kg finasteride); two diosgenin groups (50 and 100 mg/kg). The drugs were intragastricaly given in each group for consecutive 3 weeks. Another 10 rats with no testicles cut off served as negative controls and they were subcutaneously injected with 0.1 mL olive oil per day and then treated with 0.5% sodium carboxymethylcellulose. After 3-week administration, the prostate index and serum PSA level were determined, and histopathological examination was carried out. The levels of MDA, SOD and GPx in prostates were also measured. Additionally, the expression of Bcl-2, Bax and p53 was examined using Western blotting. The results showed that the prostate index and serum PSA level were significantly decreased, and the pathological changes of the prostate gland were greatly improved in diosgenin groups as compared with the model group. Elevated activities of SOD and GPx, and reduced MDA level were also observed in diosgenin-treated rats. In addition, the expression of Bcl-2 in prostates was down-regulated, whereas that of Bax and p53 was up-regulated in diosgenin-treated rats. These results indicated that diosgenin was effective in inhibiting testosterone propionate-induced prostate enlargement and may be a candidate agent for the treatment of BPH.
Assuntos
Diosgenina/uso terapêutico , Hiperplasia Prostática/tratamento farmacológico , Animais , Apoptose , Diosgenina/farmacologia , Glutationa Peroxidase/metabolismo , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo , Próstata/efeitos dos fármacos , Próstata/metabolismo , Antígeno Prostático Específico/sangue , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismoRESUMO
In this study, conventional thin-film microextraction (TFME) was endowed with magnetic by introducing superparamagnetic SiO2@Fe3O4 nanoparticles in thin-films. Novel magnetic octadecylsilane (ODS)-polyacrylonitrile (PAN) thin-films were prepared by spraying, and used for the microextraction of quetiapine and clozapine in plasma and urine samples, followed by the detection of HPLC-UV. The influencing factors on the extraction efficiency of magnetic ODS-PAN TFME, including pH, extraction time, desorption solvent, desorption time, and ion strength were investigated systematically. Under the optimal conditions, both analytes showed good linearity over ranges of 0.070-9.000µgmL(-1) and 0.012-9.000µgmL(-1) in plasma and urine samples, respectively, with correlation coefficients (R(2)) above 0.9990. Limits of detection (LODs) for quetiapine in plasma and urine samples were 0.013 and 0.003µgmL(-1), respectively. LODs for clozapine in plasma and urine samples were 0.015 and 0.003µgmL(-1), respectively. The relative standard deviations (RSDs) for quetiapine and clozapine were less than 9.23%. After the validation, the protocol was successfully applied for the determination of quetiapine and clozapine in patients' plasma and urine samples with satisfactory recoveries between 99-110%. The proposed magnetic ODS-PAN TFME was very simple, fast and easy to handle. It showed high potential as a powerful pretreatment technology for routine therapeutic drug monitoring (TDM) in plasma and urine samples.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Clozapina/sangue , Clozapina/urina , Magnetismo , Fumarato de Quetiapina/sangue , Fumarato de Quetiapina/urina , Espectrofotometria Ultravioleta/métodos , Humanos , Limite de Detecção , Microscopia Eletrônica de VarreduraRESUMO
Alzheimer's disease (AD) is one of the major neurodegenerative disorders of the elderly, which is characterized by the accumulation and deposition of amyloid-beta (Aß) peptide in human brains. Oxidative stress and neuroinflammation induced by Aß in brain are increasingly considered to be responsible for the pathogenesis of AD. The present study aimed to determine the protective effects of walnut peptides against the neurotoxicity induced by Aß25-35 in vivo. Briefly, the AD model was induced by injecting Aß25-35 into bilateral hippocampi of mice. The animals were treated with distilled water or walnut peptides (200, 400 and 800 mg/kg, p.o.) for five consecutive weeks. Spatial learning and memory abilities of mice were investigated by Morris water maze test and step-down avoidance test. To further explore the underlying mechanisms of the neuroprotectivity of walnut peptides, the activities of superoxide dismutase (SOD), glutathione (GSH), acetylcholine esterase (AChE), and the content of malondialdehyde (MDA) as well as the level of nitric oxide (NO) in the hippocampus of mice were measured by spectrophotometric method. In addition, the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), tumor necrosis factor-α (TNF-α), interleukin 1ß (IL-1ß) and IL-6 in the samples were determined using ELISA. The hippocampal expressions of inducible nitric oxide synthase (iNOS) and nuclear factor κB (NF-κB) were evaluated by Western blot analysis. The results showed that walnut peptides supplementation effectively ameliorated the cognitive deficits and memory impairment of mice. Meanwhile, our study also revealed effective restoration of levels of antioxidant enzymes as well as inflammatory mediators with supplementation of walnut peptides (400 or 800 mg/kg). All the above findings suggested that walnut peptides may have a protective effect on AD by reducing inflammatory responses and modulating antioxidant system.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Transtornos da Memória/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Peptídeos/farmacologia , Extratos Vegetais/farmacologia , Acetilcolinesterase/metabolismo , Doença de Alzheimer/etiologia , Peptídeos beta-Amiloides/toxicidade , Animais , Feminino , Glutationa/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Interleucinas/metabolismo , Juglans/química , Masculino , Malondialdeído/metabolismo , Aprendizagem em Labirinto , Transtornos da Memória/etiologia , Camundongos , NF-kappa B/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Óxido Nítrico/metabolismo , Fragmentos de Peptídeos/toxicidade , Peptídeos/uso terapêutico , Extratos Vegetais/uso terapêutico , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Two thin-film microextractions (TFME), octadecylsilane (ODS)-polyacrylonitrile (PAN)-TFME and polar enhanced phase (PEP)-PAN-TFME have been proposed for the analysis of bisphenol-A, diethylstilbestrol and 17ß-estradiol in aqueous tea extract and environmental water samples followed by high performance liquid chromatography-ultraviolet detection. Both thin-films were prepared by spraying. The influencing factors including pH, extraction time, desorption solvent, desorption volume, desorption time, ion strength and reusability were investigated. Under the optimal conditions, the two TFME methods are similar in terms of the analytical performance evaluated by standard addition method. The limits of detection for three estrogens in environmental water and aqueous tea extract matrix ranged from 1.3 to 1.6 and 2.8 to 7.1 ng mL(-1) by the two TFME methods, respectively. Both approaches were applied for the analysis of analytes in real aqueous tea extract and environmental water samples, presenting satisfactory recoveries ranged from 87.3% to 109.4% for the spiked samples.
Assuntos
Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Estrogênios/análise , Água Doce/análise , Extratos Vegetais/análise , Chá/química , Compostos Benzidrílicos/análise , Dietilestilbestrol/análise , Estradiol/análise , Análise de Alimentos , Concentração de Íons de Hidrogênio , Lagos , Concentração Osmolar , Fenóis/análise , Reprodutibilidade dos Testes , Rios , SolventesRESUMO
Novel uniform-sized magnetic molecularly imprinted polymers (MMIPs) were synthesized for selective recognition of active antitumor ingredients of kaempferol (KMF) and protoapigenone (PA) in Macrothelypteris torresiana (M. torresiana) by surface molecular imprinting technique in this study. Super paramagnetic core-shell nanoparticles (γ-MPS-SiO2@Fe3O4) were used as seeds, KMF as template molecule, acrylamide (AM) as functional monomer, and N, N'-methylene bisacrylamide (BisAM) as cross-linker. The prepared MMIPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectrum (FTIR), transmission electron microscopy (TEM) and thermo-gravimetric analysis (TGA), respectively. The recognition capacity of MMIPs was 2.436 times of non-imprinted polymers. The adsorption results based on kinetics and isotherm analysis were in accordance with the pseudo-second-order model (R (2)=0.9980) and the Langmuir adsorption model (R (2)=0.9944). The value of E (6.742 kJ/mol) calculated from the Dubinin-Radushkevich isotherm model suggested that the physical adsorption via hydrogen-bonding might be predominant. The Scatchard plot showed a single line (R (2)=0.9172) and demonstrated the homogeneous recognition sites on MMIPs for KMF. The magnetic solid phase extraction (MSPE) based on MMIPs as sorbent was established for fast and selective enrichment of KMF and its structural analogue PA from the crude extract of M. torresiana and then KMF and PA were detected by HPLC-UV. The established method showed good performance and satisfactory results for real sample analysis. It also showed the feasibility of MMIPs for selective recognition of active structural analogues from complex herbal extracts.
Assuntos
Resinas Acrílicas , Antineoplásicos Fitogênicos/isolamento & purificação , Cicloexanonas/isolamento & purificação , Gleiquênias/química , Flavonas/isolamento & purificação , Quempferóis/isolamento & purificação , Nanopartículas/química , Resinas Acrílicas/síntese química , Resinas Acrílicas/química , Antineoplásicos Fitogênicos/química , Cicloexanonas/química , Flavonas/química , Quempferóis/químicaRESUMO
P-glycoprotein (P-gp), an important efflux transporter, is encoded by the MDR1 class of genes and is a major element of the multidrug resistance (MDR) phenomenon in breast cancers. The most common approved cause of MDR in cancer tissues is the over-expression of P-gp. At present, a novel potent anti-tumor compound RY10-4 has been synthesized by our team, which has a similar structure close to protoapigenone. We chose MCF-7/ADR cells, an adriamycin (ADR) - selected human breast tumor cell line with the MDR phenotype, to study the anticancer features of this novel compound in our experiments. In cytotoxicity and apoptosis tests, it was shown that RY10-4 significantly inhibited cell growth, induced apoptosis, potentiated ADR cytotoxicity and restored chemotherapy sensitivity in the MDR cancer cells. Furthermore, our results suggested that RY10-4 reversed MDR partially by down-regulation of P-gp and MDR1 expressions in the MCF-7/ADR cell line. Besides, it is seen that RY10-4 could reduce the intracellular ATP level. Our studies give the theoretical basis for the possible clinical applications of RY10-4 alone or in combination with other chemotherapeutic drugs in the treatment of MDR tumors.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Neoplasias da Mama/metabolismo , Doxorrubicina/farmacologia , Resistência a Múltiplos Medicamentos/fisiologia , Fenótipo , Pironas/farmacologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Neoplasias da Mama/tratamento farmacológico , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/fisiologia , Doxorrubicina/uso terapêutico , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Células MCF-7 , Pironas/uso terapêuticoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Abacopteris penangiana (Hook.) Ching (AP) is a member of parathelypteris glanduligera and used in folk medicine for the treatment of blood circulation and blood stasis, edema and inflammation as recorded in the â³Chinese Materia Medicaâ³. AIM OF THE STUDY: The purpose of this study was to investigate the effects of total flavanol glycosides (TFA) from AP and its acid hydrolysate (AHT) on testosterone-induced benign prostatic hyperplasia (BPH) in rats by measuring the levels of inflammatory responses, oxidative stress and prostate cell proliferation. MATERIALS AND METHODS: BPH was induced in rats by subcutaneous injection of testosterone after castration. Seventy rats were divided into seven groups. After oral administration of AHT and TFA (100 or 200mg/kg/d) for 4 weeks, the prostate index (PI), 5a-reductase (5α-R) and dihydrotestosterone (DHT) were determined. Then the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were determined. In addition, the relative inflammatory factors, cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin 1ß (IL-1ß), interleukin 6 (IL-6), interleukin 8 (IL-8) and interleukin 17 (IL-17) were measured. Finally, the prostatic expression of nuclear transcription factor-κB (NF-κB) and phosphoinositide3-kinase (PI3K)/Akt were determined by immunohistochemistry. The prostatic expression of Bcl-2 was determined by western blot analysis. RESULTS: The results showed that AHT and TFA decreased serum DHT and 5α-R activities compared with model group, as well as the PI and histopathological examination findings. In addition, oral treatment of AHT and TFA can significantly increase the activities of SOD, GPx and CAT while the level of MDA was significantly decreased compared with the model group. Moreover, AHT and TFA remarkably decreased the levels of inflammatory cytokines in prostatic tissue. Further investigation demonstrated that AHT and TFA treatment down-regulated the protein expressions of p-Akt, NF-κB and Bcl-2. CONCLUSIONS: These results suggest that AHT and TFA have anti-BPH properties via anti-inflammatory, antioxidant and anti-proliferative effects. Hence, AP represents a potential herb for the treatment of BPH.
Assuntos
Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Hiperplasia Prostática/tratamento farmacológico , Traqueófitas/química , Animais , Antioxidantes/administração & dosagem , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Medicina Tradicional , Extratos Vegetais/administração & dosagem , Hiperplasia Prostática/patologia , Ratos , Ratos Sprague-Dawley , Testosterona/toxicidadeRESUMO
Arachniodes exilis is used as a folk medicine in China and proved to have antibacterial, anti-inflammatory, and sedative activities. In the present study, the antitumor effect of the total flavonoids of A. exilis (TFAE) against HepG2 cells was evaluated. The results showed that TFAE inhibited the growth of HepG2 cells in a dosage- and time-dependent manner. Flow cytometry and Hoechst 33342 fluorescence staining results showed that TFAE could significantly increase the apoptosis ratio of HepG2 cells, which is accompanied with increased intracellular reactive oxygen species (ROS) production and decreased mitochondrial membrane potential (ΔΨm). Western blotting indicated that TFAE downregulated the ratio of Bcl-2/Bax, increased cytochrome c release, and activated the caspases-3 and -9. Further analysis showed that TFAE stimulated the mitogen-activated protein kinase (MAPK). However, treatment with NAC (reactive oxygen species scavenger) and MAPK-specific inhibitors (SP600125 and SB203580) could reverse the changes of these apoptotic-related proteins. These results suggested that TFAE possessed potential anticancer activity in HepG2 cells through ROS-mediated mitochondrial dysfunction involving MAPK pathway.
RESUMO
Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin (1, 2 and 4 µmol/L) could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.
Assuntos
Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Diosgenina/análogos & derivados , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Diosgenina/química , Diosgenina/farmacologia , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Citometria de Fluxo , Humanos , Masculino , Estrutura Molecular , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Fatores de Tempo , Proteína X Associada a bcl-2/metabolismoRESUMO
Previous reports suggested that protoapigenone showed remarkable antitumor activities against a broad spectrum of human cancer cell lines, but had no effect on human lung adenocarcinoma A549 cell. The lack of effective remedies had necessitated the application of new therapeutic scheme. A novel compound RY10-4 which has the similar structure close to protoapigenone showed better antitumor activity. Treatment with RY10-4 inhibited the expression of pro-caspase-3, pro-caspase-9, Bcl-2 as well as phosphorylation of signal transducer and activator of transcription-3 (p-STAT3). It also reduced the expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and increases the expressions of reversion-inducing cysteine-rich protein with kazal motifs (RECK), as well as tissue inhibitor of metalloproteinase (TIMP) via inhibiting STAT3 by activating the mitogen-activated protein (MAP) kinases (the c-Jun N-terminal kinase (JNK), the p38 and extracellular signal-regulated kinase (ERK)) in A549 cells treated with RY10-4. Moreover, the cytotoxic effect of RY10-4 was induction of apoptosis in A549 cells by enhancing production of reactive oxygen species (ROS). Taken together, the observations suggested that RY10-4 had affected Bcl-2 family members, caspases, MMPs, TIMPs expressions and ROS production via inhibiting STAT3 activities through ERK and p38 pathways in A549 cells.