RESUMO
Twelve undescribed lanostane-type triterpenes, and twenty-two known triterpenes were isolated and identified from a medicinal bracket fungus Fomitopsis pinicola (Sw.) P. Karst. The structures of these compounds were determined by spectroscopic and spectrometric analyses. The antiinflammatory potential of thirty-two triterpene compounds was evaluated using neutrophils as an assay model, and pinicolasin J was the most potent inhibitor of superoxide anion generation and elastase release, with IC50 values of 1.81 ± 0.44 and 2.50 ± 0.64 µM, respectively. This study provides scientific insight into the nutritional supplement value and medicinal development of Fomitopsis pinicola.
Assuntos
Anti-Inflamatórios/farmacologia , Coriolaceae/química , Inibidores Enzimáticos/farmacologia , Carpóforos/química , Elastase Pancreática/antagonistas & inibidores , Triterpenos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Humanos , Estrutura Molecular , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Elastase Pancreática/metabolismo , Relação Estrutura-Atividade , Triterpenos/química , Triterpenos/isolamento & purificaçãoRESUMO
Four new sesquiterpenes (1-4), one new alkaloid (5), and one new benzenoid glycoside (6) were characterized from Lindera aggregata, and their structures were elucidated according to their spectrometric analytical data. Among these isolates, 3 and 4 were constructed as possessing unprecedented carbon skeletons from the natural source. Some of these purified constituents were examined for their anti-inflammatory bioactivity. Among the tested compounds, linderaggredin C (3), (+)-N-methyllaurotetanine, and (+)-isoboldine displayed the significant inhibition of superoxide anion generation in human neutrophils with IC50 values of 7.45 ± 0.74, 8.36 ± 0.11, and 5.81 ± 0.59 µM, respectively.
Assuntos
Anti-Inflamatórios/farmacologia , Lindera/química , Sesquiterpenos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Humanos , Estrutura Molecular , Neutrófilos/efeitos dos fármacos , Elastase Pancreática/metabolismo , Sesquiterpenos/química , Sesquiterpenos/isolamento & purificação , Superóxidos/metabolismoRESUMO
This study investigates the application of the paramagnetic shift reagent tris(dipivaloylmethanato)-europium(III) in NMR spectral studies of permethoxyacridone alkaloids (1-3) and pyranoacridone alkaloids (4-6). The induced chemical shifts (∆δ) of all protons were observed for the same molecule, and were compared to deduce the positions resulting from the distance nearby the Eu(dpm)3. Assignment of the H-2, H-4 and H-8 of polysubstituted acridones could be distinguished based on the least-squares method of lanthanide-induced shifts plotted against the mole ratios of Eu(dpm)3 to the substrate. The developed method is not only potentially useful for determining the planar structures of polysubstituted compounds, such as acridones, anthraquinones, xanthones, flavonoids, and phenanthrenes, but also applicable for their stereochemistry.
Assuntos
Acridonas/química , Alcaloides/química , Elementos da Série dos Lantanídeos/química , Espectroscopia de Prótons por Ressonância Magnética , Európio/químicaRESUMO
Minoxidil is clinically used to prevent hair loss. However, its effect on Ca2+ homeostasis in prostate cancer cells is unclear. This study explored the effect of minoxidil on cytosolic-free Ca2+ levels ([Ca2+]i) and cell viability in PC3 human prostate cancer cells. Minoxidil at concentrations between 200 and 800 µM evoked [Ca2+]i rises in a concentration-dependent manner. This Ca2+ signal was inhibited by 60% by removal of extracellular Ca2+. Minoxidil-induced Ca2+ influx was confirmed by Mn2+-induced quench of fura-2 fluorescence. Pre-treatment with the protein kinase C (PKC) inhibitor GF109203X, PKC activator phorbol 12-myristate 13 acetate (PMA), nifedipine and SKF96365 inhibited minoxidil-induced Ca2+ signal in Ca2+ containing medium by 60%. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-ditert-butylhydroquinone (BHQ) in Ca2+-free medium abolished minoxidil-induced [Ca2+]i rises. Conversely, treatment with minoxidil abolished BHQ-induced [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 abolished minoxidil-evoked [Ca2+]i rises. Overnight treatment with minoxidil killed cells at concentrations of 200-600 µM in a concentration-dependent fashion. Chelation of cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM (BAPTA/AM) did not prevent minoxidil's cytotoxicity. Together, in PC3 cells, minoxidil induced [Ca2+]i rises that involved Ca2+ entry through PKC-regulated store-operated Ca2+ channels and PLC-dependent Ca2+ release from the endoplasmic reticulum. Minoxidil-induced cytotoxicity in a Ca2+-independent manner.
Assuntos
Anti-Hipertensivos/farmacologia , Apoptose/efeitos dos fármacos , Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Proliferação de Células/efeitos dos fármacos , Minoxidil/farmacologia , Neoplasias da Próstata/patologia , Humanos , Masculino , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Células Tumorais CultivadasRESUMO
Fifty compounds were isolated from the fruits of Forsythia suspensa, including 13 new compounds characterized as eight new diterpenoids (1-8), three new lignans (9-11), a new iridoid (12), and a new triterpenoid (13). Their structures were established on the basis of spectroscopic and spectrometric analysis. Most of the isolated compounds were examined for their anti-inflammatory activity in vitro. The results showed that several compounds displayed significant inhibition of fMLP/CB-induced superoxide anion generation and elastase release, with IC50 values ranging from 0.6 ± 0.1 to 8.6 ± 0.8 µg/mL and from 0.8 ± 0.3 to 7.3 ± 1.1 µg/mL, respectively.
Assuntos
Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Diterpenos/isolamento & purificação , Diterpenos/farmacologia , Forsythia/química , Frutas/química , Lignanas/isolamento & purificação , Lignanas/farmacologia , Anti-Inflamatórios/química , Diterpenos/química , Humanos , Lignanas/química , Estrutura Molecular , Neutrófilos/efeitos dos fármacos , Elastase Pancreática/efeitos dos fármacos , Elastase Pancreática/metabolismo , Superóxidos/metabolismo , TaiwanRESUMO
A series of chromatographic separations performed on the ethanol extracts of the peels of Citrus grandis has led to the characterization of forty compounds, including seventeen coumarins, eight flavonoids, two triterpenoids, four benzenoids, two steroids, one lignan, one amide, and five other compounds, respectively. The chemical structures of the purified constituents were identified on the basis of spectroscopic elucidation, including 1D- and 2D-NMR, UV, IR, and mass spectrometric analysis. Most of the isolated compounds were examined for their inhibition of superoxide anion generation and elastase release by human neutrophils. Among the isolates, isomeranzin (3), 17,18-dihydroxybergamottin (12), epoxybergamottin (13), rhoifolin (19), vitexicarpin (22) and 4-hydroxybenzaldehyde (29) displayed the most significant inhibition of superoxide anion generation and elastase release with IC50 values ranged from 0.54 to 7.57 µM, and 0.43 to 4.33 µM, respectively. In addition, 7-hydroxy-8-(2'-hydroxy-3'-methylbut-3'-enyl)coumarin (8) and 17,18-dihydroxybergamottin (12) also exhibited the protection of neurons against A-mediated neurotoxicity at 50 µM.
Assuntos
Anti-Inflamatórios/farmacologia , Citrus/química , Cumarínicos/farmacologia , Flavonoides/farmacologia , Fármacos Neuroprotetores/farmacologia , Triterpenos/farmacologia , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/toxicidade , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Morte Celular/efeitos dos fármacos , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Misturas Complexas/química , Cumarínicos/química , Cumarínicos/isolamento & purificação , Flavonoides/química , Flavonoides/isolamento & purificação , Frutas/química , Humanos , Elastase de Leucócito/antagonistas & inibidores , Elastase de Leucócito/metabolismo , Lignanas/química , Lignanas/isolamento & purificação , Lignanas/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/isolamento & purificação , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Fragmentos de Peptídeos/antagonistas & inibidores , Fragmentos de Peptídeos/toxicidade , Cultura Primária de Células , Ratos , Ratos Sprague-Dawley , Esteroides/química , Esteroides/isolamento & purificação , Esteroides/farmacologia , Superóxidos/antagonistas & inibidores , Superóxidos/química , Triterpenos/química , Triterpenos/isolamento & purificação , ResíduosRESUMO
Esculetin (6,7-dihydroxycoumarin), a derivative of coumarin compound, is found in traditional medicinal herbs. It has been shown that esculetin triggers diverse cellular signal transduction pathways leading to regulation of physiology in different models. However, whether esculetin affects Ca(2+) homeostasis in breast cancer cells has not been explored. This study examined the underlying mechanism of cytotoxicity induced by esculetin and established the relationship between Ca(2+) signaling and cytotoxicity in human breast cancer cells. The results showed that esculetin induced concentration-dependent rises in the intracellular Ca(2+) concentration ([Ca(2+)]i) in ZR-75-1 (but not in MCF-7 and MDA-MB-231) human breast cancer cells. In ZR-75-1 cells, this Ca(2+) signal response was reduced by removing extracellular Ca(2+) and was inhibited by the store-operated Ca(2+) channel blocker 2-aminoethoxydiphenyl borate (2-APB). In Ca(2+)-free medium, pre-treatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) abolished esculetin-induced [Ca(2+)]i rises. Conversely, incubation with esculetin abolished TG-induced [Ca(2+)]i rises. Esculetin induced cytotoxicity that involved apoptosis, as supported by the reduction of mitochondrial membrane potential and the release of cytochrome c and the proteolytic activation of caspase-9/caspase-3, which were partially reversed by pre-chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM). Moreover, esculetin increased the percentage of cells in G2/M phase and regulated the expressions of p53, p21, CDK1, and cyclin B1. Together, in ZR-75-1 cells, esculetin induced [Ca(2+)]i rises by releasing Ca(2+) from the ER and causing Ca(2+) influx through 2-APB-sensitive store-operated Ca(2+) entry. Furthermore, esculetin activated Ca(2+)-associated mitochondrial apoptotic pathways that involved G2/M cell cycle arrest. Graphical abstract The summary of esculetin-evoked [Ca(2+)]i rises and -activated Ca(2+)-associated mitochondrial apoptotic pathways that involved cell cycle arrest. The natural coumarin derivative esculetin caused Ca(2+) influx via 2-APB-sensitive store-operated Ca(2+) entry and induced Ca(2+) release from the endoplasmic reticulum. Moreover, esculetin activated the mitochondrial pathway of apoptosis in a Ca(2+)-associated manner that involved G2/M arrest.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Umbeliferonas/farmacologia , Neoplasias da Mama/tratamento farmacológico , Cálcio , Sinalização do Cálcio , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Pontos de Checagem da Fase G1 do Ciclo Celular , Humanos , Concentração Inibidora 50 , Potencial da Membrana Mitocondrial/efeitos dos fármacos , MitocôndriasRESUMO
The effect of protriptyline on Ca2+ physiology in human hepatoma is unclear. This study explored the effect of protriptyline on [Ca2+ ]i and cytotoxicity in HepG2 human hepatoma cells. Protriptyline (50-150 µM) evoked [Ca2+ ]i rises. The Ca2+ entry was inhibited by removal of Ca2+ . Protriptyline-induced Ca2+ entry was confirmed by Mn2+ -induced quench of fura-2 fluorescence. Except nifedipine, econazole, SKF96365, GF109203X, and phorbol 12-myristate 13 acetate did not inhibit Ca2+ entry. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) inhibited 40% of protriptyline-induced response. Treatment with protriptyline abolished BHQ-induced response. Inhibition of phospholipase C (PLC) suppressed protriptyline-evoked response by 70%. At 20-40 µM, protriptyline killed cells which was not reversed by the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Together, in HepG2 cells, protriptyline induced [Ca2+ ]i rises that involved Ca2+ entry through nifedipine-sensitive Ca2+ channels and PLC-dependent Ca2+ release from endoplasmic reticulum. Protriptyline induced Ca2+ -independent cell death.
Assuntos
Antidepressivos Tricíclicos/farmacologia , Cálcio/metabolismo , Morte Celular/efeitos dos fármacos , Protriptilina/farmacologia , Cálcio/agonistas , Cátions Bivalentes , Econazol/farmacologia , Ácido Egtázico/análogos & derivados , Ácido Egtázico/farmacologia , Corantes Fluorescentes , Fura-2 , Células Hep G2 , Humanos , Hidroquinonas/farmacologia , Imidazóis/farmacologia , Indóis/farmacologia , Transporte de Íons/efeitos dos fármacos , Cinética , Maleimidas/farmacologia , Manganês/farmacologia , Nifedipino/farmacologia , Protriptilina/antagonistas & inibidores , Espectrometria de Fluorescência , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologia , Fosfolipases Tipo C/antagonistas & inibidores , Fosfolipases Tipo C/metabolismoRESUMO
Arsenic trioxide is an old drug and has been used for a long time in traditional Chinese and Western medicines. However, the cancer treatment of arsenic trioxide has heart and vascular toxicity. The cytotoxic effects of arsenic trioxide and its molecular mechanism in human umbilical mesenchymal stem cells (HUMSC) and human bone marrow-derived mesenchymal stem cells (HMSC-bm) were investigated in this study. Our results showed that arsenic trioxide significantly reduced the viability of HUMSC and HMSC-bm in a concentration- and time-dependent manner. Arsenic trioxide is able to induce apoptotic cell death in HUMSC and HMSC-bm, as shown from the results of morphological examination, flow cytometric analyses, DAPI staining and comet assay. The appearance of arsenic trioxide also led to an increase of intracellular free calcium (Ca(2+) ) concentration and the disruption of mitochondrial membrane potential (ΔΨm). The caspase-9 and caspase-3 activities were time-dependently increased in arsenic trioxide-treated HUMSC and HMSC-bm. In addition, the proteomic analysis and DNA microarray were carried out to investigate the expression level changes of genes and proteins affected by arsenic trioxide treatment in HUMSC. Our results suggest that arsenic trioxide induces a prompt induction of ER stress and mitochondria-modulated apoptosis in HUMSC and HMSC-bm. A framework was proposed for the effect of arsenic trioxide cytotoxicity by targeting ER stress.
Assuntos
Apoptose/efeitos dos fármacos , Células da Medula Óssea/efeitos dos fármacos , Estresse do Retículo Endoplasmático/fisiologia , Sangue Fetal/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Óxidos/toxicidade , Apoptose/genética , Trióxido de Arsênio , Arsenicais , Células da Medula Óssea/fisiologia , Caspase 3/metabolismo , Caspase 9/metabolismo , Células Cultivadas , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Sangue Fetal/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Células-Tronco Mesenquimais/fisiologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteoma/análise , Proteoma/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
This study explored the effect of deltamethrin, a pesticide, on intracellular free Ca²âº concentration ([Ca²âº]i) in PC3 human prostate cancer cells. Deltamethrin at concentrations between 5 µM and 20 µM evoked [Ca²âº]i rises in a concentration-dependent manner. This Ca²âº signal was inhibited by 22% by removal of extracellular Ca²âº. Nifedipine, econazole, and SKF96365 also inhibited the Ca²âº signal. Treatment with the endoplasmic reticulum Ca²âº pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) in Ca²âº-free medium nearly abolished deltamethrin-induced [Ca²âº]i rises. Treatment with deltamethrin also inhibited most of BHQ-induced [Ca²âº]i rises. Inhibition of phospholipase C (PLC) with U73122 failed to alter deltamethrin-evoked [Ca²âº]i rises. Deltamethrin killed cells at concentrations of 20-100 µM in a concentration-dependent fashion. Chelation of cytosolic Ca²âº with 1,2-bis (2-aminophenoxy) ethane-N, N, N', N'-tetraacetic acid/acetoxymethyl ester (BAPTA/AM) did not prevent deltamethrin's cytotoxicity. Together, in PC3 human prostate cancer cells, deltamethrin induced [Ca²âº]i rises that involved Ca²âº entry through store-operated Ca²âº channels and PLC-independent Ca²âº release from the endoplasmic reticulum. Deltamethrin induced cytotoxicity in a Ca²âº-independent manner.
Assuntos
Cálcio/metabolismo , Inseticidas/farmacologia , Nitrilas/farmacologia , Neoplasias da Próstata/metabolismo , Piretrinas/farmacologia , Apoptose/efeitos dos fármacos , Bloqueadores dos Canais de Cálcio/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Quelantes/farmacologia , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Inibidores Enzimáticos/farmacologia , Humanos , Masculino , Nitrilas/antagonistas & inibidores , Piretrinas/antagonistas & inibidores , Fosfolipases Tipo C/antagonistas & inibidoresRESUMO
NPC15199 is a synthesized compound that inhibits inflammation in some models. However, whether NPC15199 affects Ca²âº homeostasis in human gastric cancer is unclear. This study examined the effect of NPC15199 on cytosolic free Ca²âº concentrations ([Ca²âº]i) and viability in SCM1 human gastric cancer cells. The Ca²âº-sensitive fluorescent dye fura-2 was used to measure [Ca²âº]i. NPC15199 evoked [Ca²âº]i rises concentration-dependently. The response was reduced by removing extracellular Ca²âº. NPC15199-evoked Ca²âº entry was not inhibited by store-operated channel inhibitors (nifedipine, econazole and SKF96365) and protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate, PMA), or PKC inhibitor (GF109203X). In Ca²âº-free medium, treatment with the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) nearly abolished NPC15199-evoked [Ca²âº]i rises. Conversely, treatment with NPC15199 also nearly abolished thapsigargin or BHQ-evoked [Ca²âº]i rises. Inhibition of phospholipase C (PLC) with U73122 did not affect NPC15199-evoked [Ca²âº]i rises. NPC15199 at concentrations of 100-900 µM induced concentration-dependent, Ca²âº-independent decrease in viability. Together, in SCM1 cells, NPC15199 induced [Ca²âº]i rises that involved Ca²âº entry through PKC-insensitive non-store-operated Ca²âº channels and PLC-independent Ca²âº release from the endoplasmic reticulum. NPC15199 also induced Ca²âº-independent cell death.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , Fluorenos/uso terapêutico , Leucina/análogos & derivados , Neoplasias Gástricas/tratamento farmacológico , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Fluorenos/farmacologia , Humanos , Leucina/farmacologia , Leucina/uso terapêutico , Fosfolipases Tipo C/metabolismoRESUMO
Tricyclic antidepressants (TCA) have been clinically prescribed in the auxiliary treatment of cancer patients. Although protriptyline, a type of TCA, was used primarily in the clinical treatment of mood disorders in cancer patients, the effect of protriptyline on physiology in human osteosarcoma is unknown. This study examined the effect of protriptyline on cytosolic free Ca2+ concentrations ([Ca2+]i) and viability in MG63 human osteosarcoma cells. Protriptyline between 50 and 250 µM evoked [Ca2+]i rises concentration-dependently. Protriptyline induced influx of Mn2+, indirectly implicating Ca2+ influx. Protriptyline-evoked Ca2+ entry was inhibited by nifedipine by 20% but was not altered by econazole, SKF96365, GF109203X, and phorbol-12-myristate-13-acetate (PMA). In Ca2+-free medium, treatment with protriptyline inhibited the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin-evoked [Ca2+]i rises. Conversely, treatment with thapsigargin inhibited 45% of protriptyline-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 failed to alter protriptyline-evoked [Ca2+]i rises. Protriptyline at 50-250 µM decreased cell viability, which was not reversed by pretreatment with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, our data suggest that in MG63 cells, protriptyline induced [Ca2+]i rises by evoking Ca2+ release from the endoplasmic reticulum and other stores in a PLC-independent manner, and Ca2+ entry via a nifedipine-sensitive Ca2+ pathway. Protriptyline also caused Ca2+-independent cell death.
Assuntos
Antidepressivos Tricíclicos/toxicidade , Cálcio/metabolismo , Osteoblastos/efeitos dos fármacos , Protriptilina/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citosol/efeitos dos fármacos , Citosol/metabolismo , Relação Dose-Resposta a Droga , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Humanos , Osteoblastos/metabolismo , Osteoblastos/patologiaRESUMO
The phenolic compound 2,5-dimethylphenol is a natural product. 2,5-Dimethylphenol has been shown to affect rat hepatic and pulmonary microsomal metabolism. However, the effect of 2,5-dimethylphenol on Ca(2+ )signaling and cyotoxicity has never been explored in any culture cells. This study explored the effect of 2,5-dimethylphenol on cytosolic free Ca(2+ )levels ([Ca(2+)]i) and cell viability in PC3 human prostate cancer cells. 2,5-Dimethylphenol at concentrations between 500 µM and 1000 µM evoked [Ca(2+)]i rises in a concentration-dependent manner. This Ca(2+ )signal was inhibited by approximately half by the removal of extracellular Ca(2+). 2,5-Dimethylphenol-induced Ca(2+ )influx was confirmed by Mn(2+)-induced quench of fura-2 fluorescence. Pretreatment with the protein kinase C (PKC) inhibitor GF109203X, nifedipine or the store-operated Ca(2+ )entry inhibitors (econazole or SKF96365) inhibited 2,5-dimethylphenol-induced Ca(2+ )signal in Ca(2+)-containing medium by â¼30%. Treatment with the endoplasmic reticulum Ca(2+ )pump inhibitor thapsigargin in Ca(2+)-free medium abolished 2,5-dimethylphenol-induced [Ca(2+)]i rises. Conversely, treatment with 2,5-dimethylphenol abolished thapsigargin-induced [Ca(2+)]i rises. Inhibition of phospholipase C (PLC) with U73122 reduced 2,5-dimethylphenol-evoked [Ca(2+)]i rises by â¼80%. 2,5-Dimethylphenol killed cells at concentrations of 350-1000 µM in a concentration-dependent fashion. Chelation of cytosolic Ca(2+ )with 1,2-bis(2-aminophenoxy)ethane-N, N, N', N'-tetraacetic acid/AM (BAPTA/AM) did not prevent 2,5-dimethylphenol's cytotoxicity. Together, in PC3 cells, 2,5-dimethylphenol induced [Ca(2+)]i rises that involved Ca(2+ )entry through PKC-regulated store-operated Ca(2+ )channels and PLC-dependent Ca(2+ )release from the endoplasmic reticulum. 2,5-Dimethylphenol induced cytotoxicity in a Ca(2+)-independent manner.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Xilenos/farmacologia , Canais de Cálcio/metabolismo , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/metabolismo , Humanos , Masculino , Manganês/metabolismo , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Fatores de Tempo , Fosfolipases Tipo C/metabolismoRESUMO
Protriptyline, a tricyclic anti-depressant, is used primarily to treat the combination of symptoms of anxiety and depression. However, the effect of protriptyline on prostate caner is unknown. This study examined whether the anti-depressant protriptyline altered Ca(2+) movement and cell viability in PC3 human prostate cancer cells. The Ca(2+)-sensitive fluorescent dye fura-2 was used to measure [Ca(2+)](i). Protriptyline evoked [Ca(2+)](i) rises concentration-dependently. The response was reduced by removing extracellular Ca(2+). Protriptyline-evoked Ca(2+) entry was inhibited by store-operated channel inhibitors (nifedipine, econazole and SKF96365), protein kinase C activator (phorbol 12-myristate 13 acetate, PMA) and protein kinase C inhibitor (GF109203X). Treatment with the endoplasmic reticulum Ca(2+) pump inhibitor 2,5-di-tert-butylhydr-oquinone (BHQ) in Ca(2+)-free medium inhibited 60% of protriptyline-evoked [Ca(2+)](i) rises. Conversely, treatment with protriptyline abolished BHQ-evoked [Ca(2+)](i) rises. Inhibition of phospholipase C with U73122 suppressed 50% of protriptyline-evoked [Ca(2+)](i) rises. At concentrations of 50-70 µM, protriptyline decreased cell viability in a concentration-dependent manner; which were not reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, in PC3 cells, protriptyline evoked [Ca(2+)](i) rises by inducing phospholipase C-associated Ca(2+) release from the endoplasmic reticulum and other stores, and Ca(2+) influx via protein kinase C-sensitive store-operated Ca(2+) channels. Protriptyline caused cell death that was independent of [Ca(2+)](i) rises.
Assuntos
Apoptose/efeitos dos fármacos , Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Protriptilina/administração & dosagem , Transporte Biológico Ativo/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Masculino , Neoplasias da Próstata/tratamento farmacológicoRESUMO
Baicalein (5,6,7-trihydroxyflavone) (1) has been found to be active against a wide variety of cancer cells. However, the molecular mechanism underlying the effects of 1 on the induction of Ca(2+) movement and cytotoxicity in human breast cancer cells is unknown. This study examined the relationship between 1-induced Ca(2+) signaling and cytotoxicity in ZR-75-1 human breast cancer cells. The in vitro investigations reported herein produced the following results: (i) Compound 1 increased intracellular Ca(2+) concentration ([Ca(2+)]i) in a concentration-dependent manner. The signal was decreased by approximately 50% by removal of extracellular Ca(2+). (ii) Compound 1-triggered [Ca(2+)]i increases were significantly suppressed by store-operated Ca(2+) channel blockers 2-aminoethoxydiphenyl borate (2-APB) and the PKC inhibitor GF109203X. (iii) In Ca(2+)-free medium, compound 1-induced [Ca(2+)]i increases were also inhibited by GF109203X. Furthermore, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) or 2,5-ditert-butylhydroquinone (BHQ) abolished 1-induced [Ca(2+)]i increases. Inhibition of phospholipase C (PLC) with U73122 abolished 1-induced [Ca(2+)]i increases. (iv) Compound 1 (20-40 µM) caused cytotoxicity, increased reactive oxygen species (ROS) production, and activated caspase-9/caspase-3. Furthermore, compound 1-induced apoptosis was significantly inhibited by prechelating cytosolic Ca(2+) with BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester) or by decreasing ROS with the antioxidant NAC (N-acetylcysteine). Together, baicalein (1) induced a [Ca(2+)]i increase by inducing PLC-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via PKC-dependent, 2-APB-sensitive store-operated Ca(2+) channels. Moreover, baicalein (1) induced Ca(2+)-associated apoptosis involved ROS production in ZR-75-1 cells.
Assuntos
Cálcio/metabolismo , Flavanonas/farmacologia , Acetilcisteína/farmacologia , Animais , Apoptose/efeitos dos fármacos , Compostos de Boro , Neoplasias da Mama , Caspase 3/metabolismo , Caspase 9 , Citosol/metabolismo , Ácido Egtázico/análogos & derivados , Retículo Endoplasmático/efeitos dos fármacos , Feminino , Humanos , Indóis , Maleimidas , Estrutura Molecular , Espécies Reativas de Oxigênio/farmacologia , Tapsigargina/farmacologia , Fosfolipases Tipo CRESUMO
Eight new clausenamides, including three γ-lactams (1-3), four δ-lactams (4-7), and an amide (8), and seven known lactams, including compounds 9-11, which were purified from natural sources for the first time, were characterized from the leaves of Clausena lansium. Their structures were elucidated using spectroscopic methods, and the absolute configurations were determined using electronic circular dichroism and single-crystal X-ray diffraction analyses with Cu Kα radiation. Compound 2 (50 µM) protected 22.24% of cortical neurons against Aß25-35-induced cell death.
Assuntos
Clausena/química , Lactamas/isolamento & purificação , Lignanas/isolamento & purificação , Fármacos Neuroprotetores/isolamento & purificação , Peptídeos beta-Amiloides/farmacologia , Carbazóis , Lactamas/química , Lignanas/química , Estrutura Molecular , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Ressonância Magnética Nuclear Biomolecular , Fragmentos de Peptídeos/farmacologia , Folhas de Planta/química , Estereoisomerismo , VietnãRESUMO
The effect of the antifungal drug miconazole on Ca²âº signaling in human breast cancer cells is unknown. This study examined the effect of miconazole on cytosolic free Ca²âº concentrations ([Ca²âº]i) in ZR-75-1 human breast cancer cells. The Ca²âº-sensitive fluorescent dye fura-2 was used to measure [Ca²âº]i. Miconazole induced [Ca²âº]i rises concentration-dependently. The response was reduced by 60% by removing extracellular Ca²âº. Miconazole-induced Ca²âº entry was abolished by the protein kinase C (PKC) inhibitor GF109203X, and nifedipine, but was insensitive to econazole, SKF96365 and the protein kinase C activator phorbol 12-myristate 13 acetate (PMA). In Ca²âº-free medium, treatment with the endoplasmic reticulum Ca²âº pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) or thapsigargin (TG) greatly inhibited miconazole-evoked [Ca²âº]i rises. Conversely, treatment with miconazole abolished TG and BHQ-evoked [Ca²âº]i rises. Inhibition of phospholipase C (PLC) with U73122 abolished miconazole-induced [Ca²âº]i rises. At concentrations of 30-50 µM, micronazole killed cells in a concentration-dependent manner. This cytotoxic effect was not reversed by chelating cytosolic Ca²âº with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl (BAPTA/AM). Together, in ZR-75-1 cells, miconazole induced [Ca²âº]i rises by evoking PLC-dependent Ca²âº release from the endoplasmic reticulum, and PKC-regulated nifedipine-sensitive Ca²âº entry. Miconazole-caused cell death was not triggered by a preceding [Ca²âº]i rise.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Cálcio/metabolismo , Miconazol/farmacologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Fosfolipases Tipo C/fisiologiaRESUMO
The effect of sertraline, a selective serotonin reuptake inhibitor (SSRI), on cytosolic free Ca²âº concentrations ([Ca²âº](i)) in a rabbit corneal epithelial cell line (SIRC) is unclear. This study explored whether sertraline changed basal [Ca²âº](i) levels in suspended SIRC cells by using fura-2 as a Ca²âº-sensitive fluorescent dye. Sertraline at concentrations between 10-100 µM increased [Ca²âº](i) in a concentration-dependent manner. The Ca²âº signal was reduced by 23% by removing extracellular Ca²âº. Sertraline induced Mn²âº influx, leading to quench of fura-2 fluorescence, suggesting Ca²âº influx. This Ca²âº influx was inhibited by phospholipase A2 inhibitor aristolochic acid, but not by store-operated Ca²âº channel blockers and protein kinase C/A modulators. In Ca²âº-free medium, pretreatment with the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin, cyclopiazonic acid or 2,5-di-tert-butylhydroquinone greatly inhibited sertraline-induced Ca²âº release. Inhibition of phospholipase C with U73122 abolished sertraline-induced [Ca²âº](i) rise. At concentrations of 5-50 µM, sertraline killed cells in a concentration-dependent manner. The cytotoxic effect of 25 µM sertraline was not reversed by prechelating cytosolic Ca²âº with BAPTA/AM. Collectively, in SIRC cells, sertraline induced [Ca²âº](i) rises by causing phospholipase C-dependent Ca²âº release from the endoplasmic reticulum and Ca²âº influx via phospholipase A2-sensitive Ca²âº channels. Sertraline-caused cytotoxicity was mediated by Ca²âº-independent pathways.
Assuntos
Sinalização do Cálcio/fisiologia , Cálcio/metabolismo , Células Epiteliais/metabolismo , Epitélio Corneano/citologia , Epitélio Corneano/metabolismo , Sertralina/administração & dosagem , Animais , Sinalização do Cálcio/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Epitélio Corneano/efeitos dos fármacos , Coelhos , Inibidores Seletivos de Recaptação de Serotonina/administração & dosagemRESUMO
The effect of the antidepressant doxepin on cytosolic Ca²âº concentrations ([Ca²âº](i)) and viability in PC3 human prostate cancer cells was explored. The Ca²âº-sensitive fluorescent dye fura-2 was applied to measure [Ca²âº](i). Doxepin at concentrations of 500-1000 µM induced a [Ca²âº](i) rise in a concentration-dependent manner. The response was reduced partly by removing Ca²âº. Doxepin-evoked Ca²âº entry was suppressed by Ca²âº entry blockers (nifedipine, econazole, SK&F96365), and protein kinase C (PKC) modulators. In the absence of extracellular Ca²âº, incubation with the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) partly inhibit doxepin-induced [Ca²âº](i) rise. Incubation with doxepin nearly inhibited thapsigargin or BHQ-induced [Ca²âº](i) rise. Inhibition of phospholipase C (PLC) with U73122 failed to alter doxepin-induced [Ca²âº](i) rise. At concentrations of 200-250 µM, doxepin killed cells in a concentration-dependent manner. This cytotoxic effect was not reversed by chelating cytosolic Ca²âº with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid/acetoxy methyl (BAPTA/AM). Annexin V/PI staining data implied that doxepin (200 and 250 µM) did not induce apoptosis. Collectively, in PC3 cells, doxepin induced a [Ca²âº](i) rise by evoking PLC-independent Ca²âº release from stores including the endoplasmic reticulum and Ca²âº entry via PKC-sensitive store-operated Ca²âº channels. Doxepin caused cell death that was independent of [Ca²âº](i) rises.
Assuntos
Antidepressivos/farmacologia , Cálcio/metabolismo , Doxepina/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Humanos , Masculino , Fosfolipases Tipo C/fisiologiaRESUMO
The effect of the anti-inflammatory compound NPC-14686 on intracellular Ca²âº concentration ([Ca²âº](i)) and viability in OC2 human oral cancer cells was investigated. The Ca²âº-sensitive fluorescent probe fura-2 was used to examine [Ca²âº](i). NPC-14686 induced [Ca²âº](i) rises in a concentration-dependent fashion. The effect was reduced approximately by 10% by removing extracellular Ca²âº. NPC-14686- elicited Ca²âº signal was decreased by nifedipine, econazole, SKF96365, and GF109203X. In Ca²âº-free medium, incubation with the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) abolished NPC-14686-induced [Ca²âº](i) rises. Conversely, pretreatment with NPC-14686 abolished thapsigargin or BHQ-induced [Ca²âº](i) rises. Inhibition of phospholipase C with U73122 abolished NPC-14686-induced [Ca²âº](i) rises. At 20-100 µM, NPC-14686 inhibited cell viability, which was not reversed by chelating cytosolic Ca²âº with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid-acetoxymethyl ester (BAPTA/AM). NPC-14686 between 20 µM and 40 µM also induced apoptosis. Collectively, in OC2 cells, NPC-14686 induced [Ca²âº](i) rises by evoking phospholipase C-dependent Ca²âº release from the endoplasmic reticulum and Ca²âº entry via protein kinase C-regulated store-operated Ca²âº channels. NPC-14686 also caused Ca²âº-independent apoptosis.