RESUMO
Euphorbia dentata (Euphorbiaceae), an invasive weed, is rarely eaten by herbivorous insects and could secrete a large amount of white latex, causing a serious threat to local natural vegetation, agricultural production and human health. In order to prevent this plant from causing more negative effects on humans, it is necessary to understand and utilize the chemical relationships between the latex of E. dentata and herbivorous insects. In this study, three new norsesquiterpenes (1-3), together with seven known analogues (4-10), were isolated and identified from the latex of E. dentata. All norsesquiterpenes (1-10) showed antifeedant and growth-inhibitory effects on H. armigera with varying levels, especially compounds 1 and 2. In addition, the action mechanisms of active compounds (1-3) were revealed by detoxifying enzyme (AchE, CarE, GST and MFO) activities and corresponding molecular docking analyses. Our findings provide a new idea for the development and utilization of the latex of E. dentata, as well as a potential application of norsesquiterpenes in botanical insecticides.
Assuntos
Euphorbia , Látex , Humanos , Mecanismos de Defesa , Euphorbia/química , Helicoverpa armigera , Látex/química , Simulação de Acoplamento Molecular , AnimaisRESUMO
Three new species, Calocybe aurantiaca, C. convexa, and C. decolorata, are described based on collections made in Shenyang City, Liaoning Province, China. The main characters of C. aurantiaca are its orange-yellow sporocarps and small and smooth basidiospores. Calocybe convexa is characterized by its orange-buff pileus, very small basidiospores, and tortuous stipe, whereas C. decolorata is mainly characterized by its gills that turn blue when bruised. The sequences of nuc rDNA ITS1-5.8S-ITS2 (ITS) and the 28S D1-D5 region of the Calocybe species were analyzed, and the results indicated that the three new species belonged to the genus Calocybe and differed from other species of Calocybe. The morphological similarities of the new species to other Calocybe species and the classification system within the genus Calocybe based on molecular data are also discussed. A key is provided for the Calocybe species as reported from China in order to facilitate future studies of the genus.
Assuntos
Agaricales/classificação , Agaricales/citologia , Agaricales/genética , Agaricales/isolamento & purificação , China , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microscopia , Filogenia , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Esporos Fúngicos/citologiaRESUMO
Elucidating the coordination structures and assembling modes of atomically precise metal nanoclusters (NCs) remains a hot topic as it gives answers to the underlying mechanism of nanomaterials and bulk materials in terms of structure-property relationships. Here we report a novel silver-copper alloy NC featuring rich alkynyl-metal coordination modes and unique SbF6- assembling structures. The NC, with the composition of [Ag18Cu8(dppp)4(tBu-C6H4CîC)22](SbF6)4 (dppp = 1,3-bis(diphenylphosphino)-propane), was prepared by a stepwise synthetic approach. Single-crystal X-ray diffraction analysis revealed that such a NC featured a staircase-like Ag18Cu8 kernel, which was protected by hybrid alkynyl and dppp ligands in diverse coordination structures and multiple environments. The structural analysis also revealed the unique function of SbF6- in inducing the assembly of cluster moieties, highlighting the importance of counterions in assembling nanomolecules. The diverse coordination structures of the protective ligands with metal ions and the indispensable roles of counterions in assembling the cluster moieties have also been supported by nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESI-MS) studies, making it a model system to showcase the uniqueness of atomically precise metal NCs in illustrating the coordination chemistry of nanomaterials and bulk materials at the molecular level.
RESUMO
Atomically dispersed catalysts, with a high atomic dispersion of active sites, are efficient electrocatalysts. However, their unique catalytic sites make it challenging to improve their catalytic activity further. In this study, an atomically dispersed Fe-Pt dual-site catalyst (FePtNC) has been designed as a high-activity catalyst by modulating the electronic structure between adjacent metal sites. The FePtNC catalyst showed significantly better catalytic activity than the corresponding single-atom catalysts and metal-alloy nanocatalysts, with a half-wave potential of 0.90 V for the oxygen reduction reaction. Moreover, metal-air battery systems fabricated with the FePtNC catalyst showed peak power density values of 90.33 mW cm-2 (Al-air) and 191.83 mW cm-2 (Zn-air). By combining experiments and theoretical simulations, we demonstrate that the enhanced catalytic activity of the FePtNC catalyst can be attributed to the electronic modulation effect between adjacent metal sites. Thus, this study presents an efficient strategy for the rational design and optimization of atomically dispersed catalysts.
RESUMO
Fast and efficient detection of pollutants in the food or wastewater is an urgent need for protecting human health and ecological environment. Herein, a luminescent Zr(IV)-MOF (HBU-20) has been conveniently synthesized. It could be used as a fluorescent probe for detection of vanillin, CrO42-, and Cr2O72- in aqueous medium. All the fluorescence response time is less than 10 s and the detection limits of vanillin, CrO42- and Cr2O72- achieve 0.38 µM, 0.065 µM and 0.0089 µM, respectively. Interestingly, common anions, cations and amino acids in the solution can not affect the fluorescence detection. Meanwhile, the fluorescence detection process can be successfully implemented even under strong acid or strong alkaline conditions. Further research shows that the inner filter effect (IFE) plays a major role in the sensing process. The rapid and sensitive fluorescence responses indicate that the compound is a promising multifunctional probe for sensing toxic substance. The results can provide an important reference for the design of new fluorescent probes.
Assuntos
Benzaldeídos , Luminescência , Cátions , Fluorescência , Corantes Fluorescentes/química , HumanosRESUMO
Noble metal nanozymes have shown great promise in biomedicine; however, developing novel and high-performance noble metal nanozymes is still highly pressing and challenging. Herein, we, for the first time, prepared two-dimensional (2D) Pd@Ir bimetal nanosheets (NSs) with well-defined size and composition by a facile seed-mediated growth strategy. Enzyme-mimicked investigations find that the Pd@Ir NSs possess oxidase (OXD)-, peroxidase (POD)-, and catalase (CAT)-like multienzyme-mimetic activities. Especially, they exhibited much higher OXD- and POD-like activities than individual Pd NSs and Ir nanoparticles (NPs). The density functional theory (DFT) calculations reveal that the adsorption energy of O2 on Pd@Ir NSs is lower than that on the pure Pd NSs, which is more favorable for the conversion of O2 molecules from the triplet state (3O2) into the singlet state (1O2). Finally, based on the outstanding nanozyme activities to yield highly active singlet oxygen (1O2) and hydroxyl radicals (â¢OH) as well as excellent biosafety, the as-prepared Pd@Ir NSs were applied to treat bacteria-infected wounds, and satisfactory therapeutic outcomes were achieved. We believe that the highly efficient 2D Pd@Ir nanozyme will be an effective therapeutic reagent for various biomedical applications.
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AIM: To study the effects of 3-n-butylphthalide (NBP) on the TREK-1 channel expressed in Chinese hamster ovary (CHO) cells. METHODS: Whole-cell patch-clamp recording was used to record TREK-1 channel currents. The effects of varying doses of l-NBP on TREK-1 currents were also observed. Current-clamp recordings were performed to measure the resting membrane potential in TREK-1-transfected CHO (TREK-1/CHO) and wild-type CHO (Wt/CHO) cells. RESULTS: l-NBP (0.01-10 µmol/L) showed concentration-dependent inhibition on TREK-1 currents (IC(50)=0.06±0.03 µmol/L), with a maximum current reduction of 70% at a concentration of 10 µmol/L. l-NBP showed a more potent inhibition on TREK-1 current than d-NBP or dl-NBP. This effect was partially reversed upon washout and was not voltage-dependent. l-NBP 10 µmol/L elevated the membrane potential in TREK-1/CHO cells from -55.3 mV to -42.9 mV. However, it had no effect on the membrane potential of Wt/CHO cells. CONCLUSION: 1-NBP potently inhibited TREK-1 current and elevated the membrane potential, which may contribute to its neuroprotective activity.
Assuntos
Benzofuranos/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Canais de Potássio de Domínios Poros em Tandem/antagonistas & inibidores , Animais , Benzofuranos/administração & dosagem , Células CHO , Cricetinae , Cricetulus , Relação Dose-Resposta a Droga , Concentração Inibidora 50 , Fármacos Neuroprotetores/administração & dosagem , Técnicas de Patch-Clamp , Ratos , EstereoisomerismoRESUMO
We have reported that chymotrypsin B (CtrB) is not just a digestive enzyme but is also stored in lysosomes. Herein, we demonstrated a broad distribution of CtrB and explored the involvement of CtrB in apoptosis. Exposure of RH-35 cells to H(2)O(2) or palmitate induced the redistribution of lysosomal CtrB into the cytoplasm as a result of lysosomal membrane permeabilization (LMP). Suppression of CtrB significantly blocked apoptosis, while overexpression of CtrB sensitized apoptosis markedly. CtrB could cleave Bid under neutral conditions. In RH-35 cells with Bid silenced, apoptosis induced by CtrB protein was attenuated, suggesting that CtrB mediates apoptosis of RH-35 cells mainly through processing Bid. Our data also suggest that LMP occurs earlier than mitochondrial outer membrane permeabilization; Bid activation initiated by caspase-8 might be reinforced by CtrB in consequence of LMP, which causes a positive feedback loop leading to the accumulation of tBid, and results in lysosome- and mitochondrion-dependent apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Lisossomos/metabolismo , Proteínas/fisiologia , Ácido 4-Acetamido-4'-isotiocianatostilbeno-2,2'-dissulfônico/análogos & derivados , Apoptose/fisiologia , Caspase 8/metabolismo , Quimotripsina/metabolismo , Quimotripsina/farmacologia , Humanos , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Membranas Intracelulares/metabolismo , Lisossomos/enzimologia , Mitocôndrias/metabolismo , Mitocôndrias/fisiologia , Membranas Mitocondriais/metabolismo , Proteínas/metabolismo , Proteínas/farmacologiaRESUMO
Recently, anode materials with synergistic sodium storage mechanisms of conversion combined with alloying reactions for sodium ion batteries (SIBs) have received widespread attention due to their high theoretical capacities. In this work, through reacting with an appropriate concentration of Sb3+ ions and a simple carbonization process, hollow ZnSe/Sb2Se3 microspheres encapsulated in nitrogen-doped carbon (ZnSe/Sb2Se3@NC) are progressively synthesized based on a cation-exchange reaction, using polydopamine-coated ZnSe (ZnSe@PDA) microspheres as the precursor. Benefiting from the synergistic effects between the unique structure and composition characteristics, when serving as an anode material for SIBs, they result in higher sodium diffusion coefficients (8.7 × 10-13-3.98 × 10-9 cm2 s-1) and ultrafast pseudocapacitive sodium storage capability. Compared with ZnSe@NC and Sb2Se3@NCs exhibit, ZnSe/Sb2Se3@NC exhibits more stable capacity (438 mA h g-1 at a current of 0.5 A g-1 after 120 cycles) and superior rate performance (316 mA h g-1 at 10.0 A g-1). Our work provides a convenient method to construct high performance anodes with tunable composition and structure for energy storage.
RESUMO
Potassium channel dysfunction has been implicated in apoptosis in many pathological conditions. However, which Kv channel subunit is involved in glutamate-induced apoptosis remains unknown. In this study, the contributions of nine Kv alpha and three Kv beta subunits to glutamate-induced hippocampal neuronal apoptosis were investigated. Results showed that neuronal apoptosis was not obvious with 12 hr incubation of glutamate but increased markedly after 18 hr, which was attenuated by the Kv channel blocker TEA. Among all the Kv subunits investigated, gene and protein expression of Kv2.1 increased significantly before the appearance of neuronal apoptosis, whereas the Kv1.1 mRNA level decreased quickly, and protein expression was reduced gradually after the insult. Seven other Kv alpha subunits and three Kv beta subunits were not obviously affected over time. In addition, Kv1.1 overexpression could reduce glutamate-induced hippocampal neuronal apoptosis. Therefore, the alterations of Kv1.1 and Kv2.1 might contribute to glutamate-induced toxicity in hippocampal neurons. These findings suggest that these two Kv channel subunits may represent potential therapeutic targets for neuropathological conditions in which glutamate-induced toxicity is thought to contribute to neuronal dysfunction.
Assuntos
Apoptose/fisiologia , Ácido Glutâmico/metabolismo , Canal de Potássio Kv1.1/metabolismo , Neurônios/metabolismo , Canais de Potássio Shab/metabolismo , Animais , Western Blotting , Células Cultivadas , Fragmentação do DNA , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Marcação In Situ das Extremidades Cortadas , Neurônios/efeitos dos fármacos , Neurônios/patologia , Bloqueadores dos Canais de Potássio/farmacologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Based on multielectron conversion reactions, layered transition metal dichalcogenides are considered promising electrode materials for sodium-ion batteries, but suffer from poor cycling performance and rate capability due to their low intrinsic conductivity and severe volume variations. Here, interlayer-expanded MoSe2/phosphorus-doped carbon hybrid nanospheres coated by anatase TiO2 (denoted as MoSe2/P-C@TiO2) are prepared by a facile hydrolysis reaction, in which TiO2 coating polypyrrole-phosphomolybdic acid is utilized as a novel precursor followed by a selenization process. Benefiting from synergistic effects of MoSe2, phosphorus-doped carbon, and TiO2, the hybrid nanospheres manifest unprecedented cycling stability and ultrafast pseudocapacitive sodium storage capability. The MoSe2/P-C@TiO2 delivers decent reversible capacities of 214 mAh g-1 at 5.0 A g-1 for 8000 cycles, 154 mAh g-1 at 10.0 A g-1 for 10000 cycles, and an exceptional rate capability up to 20.0 A g-1 with a capacity of ≈175 mAh g-1 in a voltage range of 0.5-3.0 V. Coupled with a Na3V2(PO4)3@C cathode, a full cell successfully confirms a reversible capacity of 242.2 mAh g-1 at 0.5 A g-1 for 100 cycles with a coulombic efficiency over 99%.
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A novel water-soluble paclitaxel complex has been prepared by inclusion complexation with bridged bis(beta-cyclodextrin)s and characterized by means of (1)H NMR, SEM, powder X-ray diffraction patterns, TG-DTA, DSC, FT-IR, and 2D NOESY. The cyclodextrins were able to solubilize paclitaxel to levels as high as 2 mg/mL. Furthermore, the cytotoxicity of the novel complexes was assessed using a K562 leukemia cell line which indicated that drug concentrations of 10 pg/mL elicited an inhibitory effect.
Assuntos
Antineoplásicos/química , Hidrocarbonetos Aromáticos com Pontes/química , Ciclodextrinas/química , Paclitaxel/química , Poliaminas/química , Varredura Diferencial de Calorimetria , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Varredura , Modelos Moleculares , Espectroscopia de Infravermelho com Transformada de Fourier , Termogravimetria , Difração de Raios XRESUMO
Apoptosis-resistant K562 cells and apoptosis-proficient HL-60 acute myelomonocytic leukemia cells were selected to study the cell-type-specific characteristics of docetaxel. The kinetics of cytotoxicity of docetaxel showed a delayed response of K562 cells compared to HL-60 cells. After treatment with 10(-8)M docetaxel, DNA fragmentation and sub-G0/G1 cells were evident in HL-60 cells in less than 6 h, while K562 cells gradually arrested in G2/M phase of the cell cycle and appeared normal for 24 h before developing similar apoptotic changes. The delayed apoptotic changes in K562 cells were accompanied by delayed activation of caspase-3. Additionally, NADPH oxidase inhibition with diphenylene iodonium showed that reactive oxygen species (ROS) burst mediated critically in the caspase-3 activation and apoptosis in HL-60 cells but was only partially involved in those events of K562 cells. These results suggested that docetaxel exposure triggered the delayed apoptosis in K562 cells and the different ROS-dependent or independent signal pathways might account for this phenomenon. Docetaxel elicited ROS production from NADPH oxidase, which in turn triggered activation of caspase-3, leading to apoptosis in HL-60 cells. While in K562 cells, docetaxel induced apoptosis after G2/M accumulation through ROS-independent or partially dependent pathways.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Caspases/biossíntese , Espécies Reativas de Oxigênio , Taxoides/farmacologia , Caspase 3 , Docetaxel , Indução Enzimática , Células HL-60 , Humanos , Células K562 , CinéticaRESUMO
Aldehydes and acids play important roles in the fermentation inhibition of biomass hydrolysates. A series of carbonyl compounds (vanillin, syringaldehyde, 4-hydroxybenzaldehyde, pyrogallol aldehyde, and o-phthalaldehyde) were used to examine the quantitative structure-inhibitory activity relationship of carbonyl compounds on alcoholic fermentation, based on the glucose consumption rate and the final ethanol yield. It was observed that pyrogallol aldehyde and o-phthalaldehyde (5.0 mM) reduced the initial glucose consumption rate by 60 and 89%, respectively, and also decreased the final ethanol yield by 60 and 99%, respectively. Correlating the molecular descriptors to inhibition efficiency in yeast fermentation revealed a strong relationship between the energy of the lowest unoccupied molecular orbital (ELUMO) of aldehydes and their inhibitory efficiency in fermentation. On the other hand, vanillin, syringaldehyde, and 4-hydroxybenzaldehyde (5.0 mM) increased the final ethanol yields by 11, 4, and 1%, respectively. Addition of vanillin appeared to favor ethanol formation over glycerol formation and decreased the glycerol yield in yeast fermentation. Furthermore, alcohol dehydrogenase (ADH) activity dropped significantly from 3.85 to 2.72, 1.83, 0.46, and 0.11 U/mg at 6 h of fermentation at vanillin concentrations of 0, 2.5, 5.0, 10.0, and 25.0 mM correspondingly. In addition, fermentation inhibition by acetic acid and benzoic acid was pH-dependent. Addition of acetate, benzoate, and potassium chloride increased the glucose consumption rate, likely because the salts enhanced membrane permeability, thus increasing glucose consumption.
Assuntos
Aldeídos/farmacologia , Fermentação/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Álcool Desidrogenase/metabolismo , Aldeídos/química , Benzaldeídos/química , Benzaldeídos/farmacologia , Etanol/metabolismo , Glucose/metabolismo , Relação Quantitativa Estrutura-Atividade , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/enzimologia , o-Ftalaldeído/química , o-Ftalaldeído/farmacologiaRESUMO
The interactions between xylan/lignin and cellulase enzymes play a key role in the effective hydrolysis of lignocellulosic biomass. Organosolv pretreated loblolly pine (OPLP) and sweetgum (OPSG) were used to quantitatively elucidate the distinct roles of residual xylan and lignin on enzymatic hydrolysis, based on the initial hydrolysis rates and the final hydrolysis yields. The initial hydrolysis rates of OPLP and OPSG were 1.45 (glucose) and 1.19 g/L/h (glucose), respectively, under the enzyme loading of 20 FPU/g glucan. The final glucan hydrolysis yields of OPLP and OPSG at 72 h were 76.4 and 98.9%, respectively. By correlating the amount of residual lignin and xylan to the initial hydrolysis rate and the final hydrolysis yield in OPLP and OPSG, a more accurate fundamental understanding of the roles of xylan and lignin in limiting the enzymatic hydrolysis has been developed. The higher amount of residual xylan (9.7%) in OPSG resulted in lower initial hydrolysis rate (1.19 g/L/h). The higher amount of residual lignin in OPLP (18.6%) resulted in lower final hydrolysis yield of glucan (76.4%). In addition, we observed in the simultaneous saccharification and fermentation (SSF) that ethyl xyloside was produced by the enzymatic catalysis of xylose/xylan and ethanol.
Assuntos
Lignina/química , Liquidambar/química , Pinus taeda/química , Xilanos/química , Celulase/metabolismo , Etanol , Fermentação , Glucanos/química , Hidrólise , Poligalacturonase/metabolismo , Saccharomyces cerevisiae/enzimologia , Xilose/químicaRESUMO
Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB), the pre-drug of 3-n-butylphthalide (dl-NBP), had the significantly therapeutic effect on the acute cerebral ischemia. The present study was to investigate the effect of dl-PHPB on the cognitive deficits induced by chronic cerebral hypoperfusion. Rats were orally administered three doses of dl-PHPB (13, 39 and 129mg/kg), dl-NBP 100mg/kg, and piracetam 600mg/kg daily for 21 days after the bilateral permanent occlusion of the common carotid arteries. The results showed that dl-PHPB, dl-NBP and piracetam significantly improved the spatial learning and memory deficits, and the effectiveness of dl-PHPB at dose of 39mg/kg was strongest. Meanwhile, the drugs decreased superoxide dismutase activity, reduced lipid peroxide and astrocyte activation in the cortex of the hypoperfused rats. Furthermore, dl-PHPB markedly reduced white matter rarefaction. The results indicated that preventing neuropathological alterations, inhibiting oxidative damage and inflammatory reaction might contribute to the improvement of dl-PHPB on hypoperfusion-induced cognitive deficits. Therefore, dl-PHPB has therapeutic potential for the treatment of dementia caused by decrease of cerebral blood flow.
Assuntos
Benzoatos/farmacologia , Encéfalo/efeitos dos fármacos , Deficiências da Aprendizagem/tratamento farmacológico , Transtornos da Memória/tratamento farmacológico , Pentanos/farmacologia , Animais , Arteriopatias Oclusivas/complicações , Astrócitos/efeitos dos fármacos , Astrócitos/fisiologia , Encéfalo/irrigação sanguínea , Encéfalo/patologia , Doenças das Artérias Carótidas/complicações , Doença Crônica , Deficiências da Aprendizagem/etiologia , Deficiências da Aprendizagem/psicologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/etiologia , Transtornos da Memória/psicologia , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
In this study, we investigated the antitumor effect of a tumor vaccine prepared from H22 hepatocarcinoma cells induced by cartilage polysaccharide. We found out there were specific antigens which combined with antigen-specific antibodies from immune murine serum. Results of western blot analysis showed that about 36 kDa make specific antibodies appeared specific antibodies in antiserum of immune mice, whereas the best immune effects became visible at the induction time of 48 h. Analyses of 2-dimensional electrophoresis identified the specific antigen was annexin A2, which was a glycosylated protein that contained a glycosylation site, closely related to oncogenesis, cancer development, invasion and metastasis. Proteomics indicated that both quantity and conformation of annexin A2 were changed after induced by cartilage polysaccharide. Lastly, we found there was a major increase of annexin A2 mRNA on H22 cells induced by cartilage polysaccharide. In summary, our data suggested that annexin A2, a specific antigen played a key role in antitumor immune response and activating the immune system. It would be a potential type of tumor vaccine which provided new ideas for tumor immunoprophylaxis.
Assuntos
Anexina A2/metabolismo , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/prevenção & controle , Cartilagem/química , Neoplasias Hepáticas Experimentais/imunologia , Neoplasias Hepáticas Experimentais/prevenção & controle , Polissacarídeos/farmacologia , Sequência de Aminoácidos , Animais , Anexina A2/imunologia , Apoptose/efeitos dos fármacos , Western Blotting , Vacinas Anticâncer/uso terapêutico , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Eletroforese em Gel Bidimensional , Feminino , Imunização , Neoplasias Hepáticas Experimentais/metabolismo , Masculino , Camundongos , Dados de Sequência Molecular , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Regulação para CimaRESUMO
We investigated the antitumor effect of a short-chain polysaccharide (PS) extracted from porcine cartilage both in L1210 leukemic cells in vitro and in mice bearing L1210 ascites tumors. Our results show that treatment with PS resulted in a significant (P < 0.01) inhibition of cell proliferation and caused apoptotic death in L1210 cells. We also found that PS increased the ratio of Bax/Bcl-2 and activated caspase-9 and caspase-3 but not caspase-8. These results point to the activation of the mitochondrial apoptotic pathway in response to the treatment of L1210 cells with PS. Our results suggest that PS has the potential to provide significant natural defence against leukemias and lymphomas.
Assuntos
Apoptose/efeitos dos fármacos , Cartilagem/química , Proliferação de Células/efeitos dos fármacos , Leucemia L1210/tratamento farmacológico , Polissacarídeos/farmacologia , Animais , Proteínas Reguladoras de Apoptose , Western Blotting , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Fragmentação do DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Feminino , Imunofluorescência , Leucemia L1210/metabolismo , Leucemia L1210/patologia , Camundongos , Camundongos Endogâmicos DBA , Polissacarídeos/química , Proteínas , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Suínos , Fatores de Tempo , Proteína X Associada a bcl-2/metabolismoRESUMO
The hierarchy of events accompanying induction of apoptosis by the microtubule inhibitor docetaxel was investigated in HL-60 human leukemia cells. Treatment of HL-60 cells with docetaxel resulted in the production of reactive oxygen species (ROS), activation of caspase-3 (-like) protease, c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) activation, bcl-2 phosphorylation and apoptosis. Docetaxel elicited ROS production from NADPH oxidase as demonstrated by specific oxidase inhibitor diphenylene iodonium (DPI). ROS mediated the caspase-3 activation and apoptosis in HL-60 cells. The caspase inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO) effectively inhibited JNK/SAPK activation, bcl-2 phosphorylation and partially attenuated the ROS production induced by docetaxel. Docetaxel-induced bcl-2 phosphorylation was completely blocked by expression of dominant negative JNK or the JNK/SAPK inhibitor SP600125. Overexpression of bcl-2 partially prevented docetaxel-mediated ROS production and subsequent caspase-3 activation, thereby inhibiting apoptotic cell death. It is thus conferred that such sequent events as ROS production, caspase activation, JNK/SAPK activation, bcl-2 phosphorylation and the further generation of ROS should be parts of an amplification loop to increase caspase activity, thereby facilitating the apoptotic cell death process.
Assuntos
Caspases/metabolismo , Taxoides/farmacologia , Apoptose/efeitos dos fármacos , Caspase 3 , Inibidores de Caspase , Células Cultivadas , Citosol , Docetaxel , Ativação Enzimática/efeitos dos fármacos , Células HL-60 , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Modelos Biológicos , Fosforilação , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , TransfecçãoRESUMO
The inclusion complexation behavior of paclitaxel with a series of oligo(ethylenediamino) bridged bis(beta-cyclodextrin)s possessing bridge chains in different length (1-4) has been investigated in order to improve the water solubility of paclitaxel. It is found that only the long-tethered bis(beta-cyclodextrin)s 1 and 2 can form the inclusion complexes with paclitaxel, which are characterized by NMR, SEM, XRD, FT-IR, TG-DTA, DSC, and microcalorimetry technology. The results obtained show that bis(beta-cyclodextrin)s 1 and 2 are able to solubilize paclitaxel to high levels up to 2 and 0.9 mg/mL, respectively. The high complex stability of bis(beta-cyclodextrin) 1 and paclitaxel is discussed from thermodynamic viewpoint. Furthermore, the cytotoxicity of these complexes assessed using a human erythroleukemia K562 cell line indicates that the IC(50) value of 1/paclitaxel complex is 6.0 x 10(-10) mol/dm(3) (calculated as paclitaxel molar concentration), which means that the antitumor activity of 1/paclitaxel complex is better than that of parent paclitaxel (IC(50) value 9.8 x 10(-10) mol/dm(3)). This high antitumor activity, along with the satisfactory water solubility and high thermal stability of the 1/paclitaxel complex, will be potentially useful for its clinical application as a highly effective antitumor drug.