RESUMO
In this research, we evaluated the aphicidal effect of the ethanolic extract of stems and bark of Ficus petiolaris Kunth (Moraceae), in laboratory bioassays in an artificial diet against apterous adult females of Melanaphis sacchari Zehntner (Hemiptera: Aphididae). The extract was evaluated at different concentrations (500, 1,000, 1,500, 2,000, and 2,500 ppm), and the highest percentage of mortality (82%) was found at 2,500 ppm after 72 h. The positive control imidacloprid (Confial®) at 1% eliminated 100% of the aphids, and the negative control (artificial diet) only presented mortality of 4%. The chemical fractionation of the stem and bark extract of F. petiolaris yielded five fractions of FpR1-5, which were each evaluated at 250, 500, 750, and 1,000 ppm. FpR2 had the strongest aphicidal effect, with 89% mortality at 72 h at 1,000 ppm. The pure xanthotoxin compound extracted from this fraction was even more effective, with 91% aphid mortality after 72 h at 100 ppm. The lethal concentration (LC50) of xanthotoxin was 58.7 ppm (72 h). Our results indicate that the extract of F. petiolaris showed toxic activity against this aphid, and its xanthotoxin compound showed strong aphicidal activity at low concentrations.
Assuntos
Afídeos , Ficus , Sorghum , Animais , Feminino , Metoxaleno , Extratos Vegetais/farmacologiaRESUMO
The treatment of vitiligo includes the combination of psoralens and ultraviolet type A exposure. Psoralens belong to a group of natural furanocoumarins that cause the skin to become sensitive temporarily to ultraviolet type A. The aim of this study was to develop a physiologically based pharmacokinetic model of 5-MOP from Brosimum gaudichaudii to support psoralen and ultraviolet type A therapy. A study of rats was used to establish and validate rat tissue distribution. The same chemical-specific parameters used in the rat model were also employed in the human model to project human pharmacokinetics. The highest exposures in the rats were in the brain and skin. Following a single dose of 1.2 mg/kg 5-MOP in humans, the model predicted a maximum concentration of 20 ng/mL and an area under the curve of 125 ng.h/mL, matching clinical results. The half-maximum melanogenesis concentrations in B16F10 cells were 29.5, 18.5, 11.5, and 6.5 ng/mL for synthetic 5-MOP, synthetic 5-MOP with ultraviolet type A, B. gaudichaudii alone, and B. gaudichaudii plus ultraviolet type A, respectively. Physiologically based pharmacokinetic model prediction in humans supported a once-every-two-day regimen for optimal melanin production. This type of framework can be applied to support strategies for dose selection and to investigate the impact of drugs on melanocyte recovery.
Assuntos
Furocumarinas , Moraceae , 5-Metoxipsoraleno , Animais , Humanos , Metoxaleno , Fitoterapia , RatosRESUMO
Eight alkaloids (1â»8) were isolated from Ruta graveolens, and their herbicide activities were evaluated through in vitro, semivivo, and in vivo assays. The most relevant results were observed for Compounds 5 and 6â»8 at 150 µM, which decreased dry biomass by 20% and 23%, respectively. These are significant results since they presented similar values with the positive control, commercial herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Based on the performed assays, Compound 5 (graveoline) is classified as an electron-transport inhibitor during the light phase of photosynthesis, as well as a plant-growth regulator. On the other hand, Compounds 6â»8 inhibited electron and energy transfers, and are also plant-growth inhibitors. These phytotoxic behaviors based on acridone and quinolone alkaloids may serve as a valuable tool in the further development of a new class of herbicides since natural products represent an interesting alternative to replace commercial herbicides, potentially due their low toxicity.
Assuntos
Alcaloides/isolamento & purificação , Metoxaleno/análogos & derivados , Fotossíntese/efeitos dos fármacos , Ruta/química , Alcaloides/química , Alcaloides/farmacologia , Biomassa , Diurona/farmacologia , Transporte de Elétrons , Herbicidas/farmacologia , Metoxaleno/química , Metoxaleno/isolamento & purificação , Metoxaleno/farmacologiaRESUMO
Oral therapy with 8-methoxypsoralen (8-MOP) may cause major side effects, whereas the topical treatment might not be much effective due to the low penetration induced by typical formulations. Therefore, the objectives of this work are the development and characterization of a nanoemulsion (NE) containing 8-MOP together with an ex vivo permeation study, monitored by a validated HPLC-Fluo method, to determine the amount of drug retained in viable skin (epidermis (E) and dermis (D)) and in stratum corneum (SC). The optimized conditions for NE formulation were achieved by full factorial designs (25 and 32): 60â¯s and 60% of ultrasound time and potency, respectively; 10â¯mL of final volume; 2% v/v of oil phase (clove essential oil); and 10% m/v of Poloxamer 407. The NE showed mean droplet diameter of 24.98⯱â¯0.49â¯nm, polydispersity index (PDI) of 0.091⯱â¯0.23, pH values of 6.54⯱â¯0.06, refractive index of 1.3525⯱â¯0.0001 and apparent viscosity of 51.15⯱â¯3.66â¯mPa at 20⯰C. Droplets with nanospherical diameters were also observed by transmission electron microscopy (TEM). Ex vivo permeation study showed that 8.5% of the applied 8-MOP dose permeated through the biological membranes, with flux (J) of 1.35⯵gâ¯cm-2â¯h-1. The drug retention in Eâ¯+â¯D and in SC was 10.15⯱â¯1.36 and 1.95⯱â¯0.71⯵gâ¯cm-2, respectively. Retention in viable skin induced by the NE was almost two-fold higher than a compounded cream (5.04⯱â¯0.30⯵gâ¯cm-2). These results suggested that the developed NE is a promising alternative for 8-MOP topical therapy when compared to commercial formulations.
Assuntos
Metoxaleno/administração & dosagem , Nanopartículas/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Pele/metabolismo , Administração Cutânea , Animais , Óleo de Cravo/administração & dosagem , Óleo de Cravo/química , Óleo de Cravo/farmacocinética , Composição de Medicamentos , Sistemas de Liberação de Medicamentos , Estabilidade de Medicamentos , Emulsões , Metoxaleno/química , Metoxaleno/farmacocinética , Nanopartículas/química , Permeabilidade , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Poloxâmero/administração & dosagem , Poloxâmero/química , Poloxâmero/farmacocinética , Absorção Cutânea , Solubilidade , SuínosRESUMO
Pigmented purpuric dermatoses are chronic vascular inflammatory conditions characterized by the presence of pigmented macules. Among its different presentations, lichen aureus is distinguished by the lichenoid conformation of its plaques and the predilection for lower limb involvement. Its segmented form is rare and difficult to control, especially in cases of symptomatic lesions. We report a rare case of segmental lichen aureus with six years of evolution associated with light itching. We also discuss the main therapeutic approaches to control the disease.
Assuntos
Erupções Liquenoides/patologia , Betametasona/uso terapêutico , Feminino , Glucocorticoides/uso terapêutico , Humanos , Erupções Liquenoides/terapia , Metoxaleno/uso terapêutico , Pessoa de Meia-Idade , Fármacos Fotossensibilizantes/uso terapêutico , Luz SolarRESUMO
Abstract: Pigmented purpuric dermatoses are chronic vascular inflammatory conditions characterized by the presence of pigmented macules. Among its different presentations, lichen aureus is distinguished by the lichenoid conformation of its plaques and the predilection for lower limb involvement. Its segmented form is rare and difficult to control, especially in cases of symptomatic lesions. We report a rare case of segmental lichen aureus with six years of evolution associated with light itching. We also discuss the main therapeutic approaches to control the disease.
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Erupções Liquenoides/patologia , Luz Solar , Betametasona/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Erupções Liquenoides/terapia , Glucocorticoides/uso terapêutico , Metoxaleno/uso terapêuticoRESUMO
The increasing tolerance to currently-used fungicides is a major problem both in clinical and agricultural areas leading to an urgent need for the development of novel antifungal strategies. This study investigated the in vitro antimicrobial photo treatment (APT) of conidia of the plant-pathogenic fungus Colletotrichum acutatum and the ascomycete Aspergillus nidulans with the furocoumarins 8-methoxypsoralen (8-MOP) and isopimpinellin, and a mixture of two coumarins (7-methoxy coumarin and citropten). Subcellular localization of the photosensitizer 8-MOP was also determined in C. acutatum conidia. Additionally, the effects of APT on the leaves of the plant host Citrus sinensis were determined. APT with 8-MOP (50µM) led to a reduction of approximately 4 logs in the survival of the conidia of both species, and the mixture of the two coumarins (12.5mgL(-1)) resulted in a reduction of approximately 4 logs for A. nidulans and 3 logs for C. acutatum. Isopimpinellin (50µM) displayed a reduction of 4 logs for A. nidulans but less than 2 logs for C. acutatum. Washing the conidia to remove unbound photosensitizers before light exposure reduced the photodynamic inactivation of C. acutatum both with 8-MOP and the mixture of the two coumarins. The reduction was smaller for A. nidulans. 8-MOP spread throughout the cytoplasm and accumulated in structures such as lipid bodies of C. acutatum conidia. No damage to orange tree leaves was observed after APT with any of the photosensitizers.
Assuntos
Antifúngicos/farmacologia , Aspergillus nidulans/efeitos dos fármacos , Colletotrichum/efeitos dos fármacos , Cumarínicos/farmacologia , Furocumarinas/farmacologia , Metoxaleno/farmacologia , Citrus/química , Citrus/microbiologia , Colletotrichum/patogenicidade , Cumarínicos/química , Cumarínicos/isolamento & purificação , Furocumarinas/isolamento & purificação , Metoxaleno/isolamento & purificação , Estrutura Molecular , Fármacos Fotossensibilizantes/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/microbiologia , Esporos Fúngicos/efeitos dos fármacos , Luz SolarRESUMO
By isolating putative binding partners through the two-hybrid system (THS) we further extended the characterization of the specific interstrand cross-link (ICL) repair gene PSO2 of Saccharomyces cerevisiae. Nine fusion protein products were isolated for Pso2p using THS, among them the Sak1 kinase, which interacted with the C-terminal ß-CASP domain of Pso2p. Comparison of mutagen-sensitivity phenotypes of pso2Δ, sak1Δ and pso2Δsak1Δ disruptants revealed that SAK1 is necessary for complete WT-like repair. The epistatic interaction of both mutant alleles suggests that Sak1p and Pso2p act in the same pathway of controlling sensitivity to DNA-damaging agents. We also observed that Pso2p is phosphorylated by Sak1 kinase in vitro and co-immunoprecipitates with Sak1p after 8-MOP+UVA treatment. Survival data after treatment of pso2Δ, yku70Δ and yku70Δpso2Δ with nitrogen mustard, PSO2 and SAK1 with YKU70 or DNL4 single-, double- and triple mutants with 8-MOP+UVA indicated that ICL repair is independent of YKu70p and DNL4p in S. cerevisiae. Furthermore, a non-epistatic interaction was observed between MRE11, PSO2 and SAK1 genes after ICL induction, indicating that their encoded proteins act on the same substrate, but in distinct repair pathways. In contrast, an epistatic interaction was observed for PSO2 and RAD52, PSO2 and RAD50, PSO2 and XRS2 genes in 8-MOP+UVA treated exponentially growing cells.
Assuntos
Dano ao DNA , Endodesoxirribonucleases/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas de Saccharomyces cerevisiae/genética , Reagentes de Ligações Cruzadas/farmacologia , Proteínas de Ligação a DNA/genética , Metoxaleno/farmacologia , Proteína Rad52 de Recombinação e Reparo de DNA/genética , Saccharomyces cerevisiae , Técnicas do Sistema de Duplo-Híbrido , Raios UltravioletaRESUMO
Catecóis são derivados do benzeno, podendo apresentar citotoxicidade, que pode constituir um modelo experimental útil para o desenvolvimento de novos fármacos. No bioma brasileiro inúmeras plantas produzem metabólitos com atividades diversas, como antioxidantes, ou inibidores do crescimento celular. No Brasil, as neoplasias são a segunda causa de óbito, especialmente aquelas derivadas do sistema nervoso, aumentando o interesse por novos antineoplásicos e agentes neuroprotetores. Este trabalho caracteriza efeitos citotóxicos do 1,2-dihidroxibenzeno (CAT) e discretamina (DSC) em células do sistema nervoso in vitro. Determinou-se a EC50 de CAT e DSC usando brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium (MTT), investigou-se sua auto-oxidação por espectrofotometria, avaliou-se mudanças morfológicas e condensação/fragmentação nuclear por microscopia. Avaliou-se a proteção de DSC e 8-metoxipsoraleno (8-MOP) contra a citotoxicidade do CAT. O padrão de morte celular foi analisado por citometria de fluxo. A espoliação de glutation reduzido (GSH) foi analisada usando monoclorobimano. A toxicidade do CAT para células SH-SY5Y e C6 depende da dose e associa-se à formação de quinonas. Houve mudanças morfológicas, condensação/fragmentação da cromatina e morte apoptótica, não relacionada à espoliação de GSH. DSC não foi tóxica para células SH-SY5Y, porém protegeu contra os efeitos do CAT em baixas concentrações. DSC mostrou-se citotóxica para células de glioma (GL-15 e C6) e potencializou o CAT. Pré-tratamento por 30 minutos com DSC protegeu contra a ação do CAT após 72 horas. 8-MOP potencializou os efeitos do CAT, não revertendo seus efeitos na viabilidade celular, morfologia celular, condensação/fragmentação nuclear, e espoliação de GSH. Esses resultados caracterizam um modelo de citotoxicidade que pode ser aplicado no desenvolvimento de novos agentes farmacológicos. Estudos complementares são necessários para elucidar a proteção da DSC.
Catechols are benzene derivatives, which may exhibit cytotoxic activity that can be employed to develop new drugs. Plants are important sources of metabolites with pharmacological activities such as antioxidants, or cell growth inhibitors. In Brazil, cancer is the second leading cause of death, especially those derived from the nervous system, which increase the interest for new antineoplastic and neuroprotective drugs. The cytotoxic effects promoted by 1,2-dihydroxybenzene (CAT) and discretamine (DSC) in nervous system cells were characterized in vitro. The protective effects of DSC and 8-methoxypsoralen (8-MOP) against CAT-induced cytotoxicity were also evaluated. CAT and DSC EC50 was determined by using 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT). CAT auto-oxidation was investigated by spectrophotometry. Morphological changes and nuclear condensation/ fragmentation were evaluated by microscopy. The pattern of cell death was obtained by flow cytometry. Reduced glutathione (GSH) depletion was analyzed by using monochlorobimane. CAT induced a dose-dependent toxicity to SH-SY5Y and C6 cells, associated with reactive quinones formation. It also induced morphological changes, nuclear condensation/fragmentation, and apoptotic death not caused by GSH depletion. DSC was not toxic to SH-SY5Y cells, but protected against CAT effects at low concentrations. DSC was be cytotoxic to glioma cells (GL-15 and C6) and potentiated CAT effects. However, pretreatment for 30 minutes with DSC protected them against CAT after 72 hours. 8-MOP also potentiated CAT effects instead to protect cells. These results characterize an experimental model useful for studies searching new pharmacological agents. However, further studies are needed to elucidate the DSC protective effects.
Assuntos
Humanos , /administração & dosagem , /análise , /farmacologia , /uso terapêutico , Metoxaleno/análise , Metoxaleno/farmacologia , Metoxaleno/uso terapêutico , Neoplasias/patologia , Neoplasias/prevenção & controleRESUMO
Two adolescent females and a girl, all with clinically diagnosed vitiligo, were treated with 0.2% 8-methoxypsoralen cream followed by exposure to solar ultraviolet light. One year later, they developed hypopigmented and achromic spots on the areas affected by the vitiligo. Biopsy of skin tissue taken from one of these cases showed a marked reduction in melanin. Clinical and histological findings point to a diagnosis of leukoderma punctata.
Assuntos
Metoxaleno/efeitos adversos , Terapia PUVA/efeitos adversos , Fármacos Fotossensibilizantes/efeitos adversos , Transtornos da Pigmentação/etiologia , Adolescente , Criança , Feminino , Humanos , Metoxaleno/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Transtornos da Pigmentação/patologia , Vitiligo/tratamento farmacológicoRESUMO
Duas adolescentes e uma menina com vitiligo clinicamente diagnosticado foram tratadas com 8-metoxipsoraleno a 0,2 por cento em creme Lanette com subsequente exposição solar. Um ano após, apresentaram máculas acrômicas na área do vitiligo. A biópsia de pele em um dos casos revelou melanócitos com escassa pigmentação melânica. Os achados clínicos e histológicos sugerem o diagnóstico de leucodermia punctata.
Two adolescent females and a girl, all with clinically diagnosed vitiligo, were treated with 0.2 percent 8-methoxypsoralen cream followed by exposure to solar ultraviolet light. One year later, they developed hypopigmented and achromic spots on the areas affected by the vitiligo. Biopsy of skin tissue taken from one of these cases showed a marked reduction in melanin. Clinical and histological findings point to a diagnosis of leukoderma punctata.
Assuntos
Adolescente , Criança , Feminino , Humanos , Metoxaleno/efeitos adversos , Terapia PUVA/efeitos adversos , Fármacos Fotossensibilizantes/efeitos adversos , Transtornos da Pigmentação/etiologia , Metoxaleno/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Transtornos da Pigmentação/patologia , Vitiligo/tratamento farmacológicoRESUMO
PURPOSE: To discuss the contribution of psoralen and bergapten metabolites on psoralens toxicity. METHODS: Computational chemistry prediction of metabolic reactions and toxicophoric groups based on the expert systems Derek and Meteor. RESULTS: a total of 15 metabolites were suggested for both psoralen and bergapten based on phase 1 and 2 biotransformations until the 3rd generation. Five toxicophoric substructures were shared among psoralen, bergapten and their corresponding metabolites; one toxicophoric marker (resorcinol) was only identified in bergapten and its biotransformation products. CONCLUSION: Although the toxic effects of psoralens are well known and documented, there is little information concerning the role of their metabolites in this process. We believe this work add to the knowledge of which molecular substructures are relevant to the process of metabolism and toxicity induction, thus guiding the search and development of more effective and less toxic drugs to treat vitiligo.
Assuntos
Sistemas Inteligentes , Ficusina/metabolismo , Ficusina/toxicidade , Metoxaleno/análogos & derivados , Fármacos Fotossensibilizantes/metabolismo , Fármacos Fotossensibilizantes/toxicidade , 5-Metoxipsoraleno , Animais , Biotransformação , Biologia Computacional , Simulação por Computador , Descoberta de Drogas/métodos , Humanos , Metoxaleno/metabolismo , Metoxaleno/toxicidade , Vitiligo/tratamento farmacológicoRESUMO
CYP2A enzymes are responsible for nicotine metabolism and for activating tobacco-related carcinogens. Inhibition of CYP2A is a promising approach in chemoprevention, which could lead to a decrease in cigarette consumption and to a reduction in tobacco-related cancer risk. 8-Methoxypsoralen (8-MOP) is a mechanism-based inhibitor of human CYP2A6 and CYP2A13. 8-MOP is also an inhibitor of Cyp2a5, but the mode of this inhibition is unknown. There is no published data on the inhibition of CYP2A3 by 8-MOP. The objective of this work was to investigate the characteristics of 8-MOP inhibition on mouse hepatic Cyp2a5 and rat nasal CYP2A3, in order to determine the best experimental model for chemoprevention studies using 8-MOP. The results show that 8-MOP inhibits CYP2a5 through three different mechanisms: competitive, non-competitive (K(iu)=1.7 microM), and mechanism-based (K(inactivation) of 0.17 min(-1)). By contrast, 8-MOP was able to inhibit CYP2A3-mediated coumarin 7-hydroxylase only in a non-competitive way (K(iu)=0.22 microM). In conclusion, we showed that 8-MOP inhibits Cyp2a5 and CYP2A3 through different mechanisms.
Assuntos
Hidrocarboneto de Aril Hidroxilases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Metoxaleno/farmacologia , Oxigenases de Função Mista/antagonistas & inibidores , Animais , Citocromo P-450 CYP2A6 , Família 2 do Citocromo P450 , Cinética , Masculino , Camundongos , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Ratos , Ratos WistarRESUMO
A method for sample preparation and analysis by high-performance liquid chromatography with UV detection (HPLC-UV) was developed for analysis of psoralen, bergapten and 5-[3-(4,5-dihydro-5,5-dimethyl-4-oxo-2-furanyl)-butoxy]-7H-furo[3-2-g][1]benzopyran-7-one in capsules and tablets employed in Brazil for certain illnesses. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. Analytical curves for furanocoumarins were linear in the range of 1.0-50.0 microg/ml. The recoveries of the furanocoumarins in the products analyzed were 97.3-99.5%, and the percent coefficient of variation for the quantitative analysis of the furanocoumarins in the analyses was under 5%. For inter-equipment study gas chromatography (GC) was employed.
Assuntos
Benzopiranos/análise , Ficusina/análise , Furocumarinas/análise , Metoxaleno/análogos & derivados , Fármacos Fotossensibilizantes/análise , 5-Metoxipsoraleno , Cápsulas/química , Cromatografia Líquida de Alta Pressão , Metoxaleno/análise , Preparações de Plantas/análise , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrofotometria Ultravioleta , Comprimidos/químicaRESUMO
X-ray diffraction studies carried out on single crystals of the monomeric, viz. 5,6-dimethoxy-2H-furo[2,3-h][1]benzopyran-2-one, C(13)H(10)O(5), and dimeric, viz. 5,5',6,6'-tetramethoxy-3,3',4,4'-tetrahydro-2H,2'H-3,3':4,4'-bi(furo[2,3-h][1]benzopyran)-2,2'-dione, C(26)H(20)O(10), forms of pimpinellin have revealed that, following cyclodimerization, the carbonyl groups are head-to-head with respect to one another. In the monomer, the heterocyclic ring is planar, but it exhibits a twisted-boat conformation in the dimer. Both the monomer and the dimer interact through C-H...O interactions.
Assuntos
Magnoliopsida/química , Metoxaleno/análogos & derivados , Metoxaleno/química , Raízes de Plantas/química , Dimerização , Modelos Moleculares , Conformação MolecularRESUMO
Cutaneous malignant melanoma is a very serious form of skin cancer that arises from melanocytes. Currently there is no effective treatment for metastatic melanoma so intense clinical trials are evaluating new drugs for this human malignancy. Psoralens are a group of compounds that bind to DNA in rapidly dividing cells and with ultraviolet light in the A band (UVA) cause DNA crosslinking, thereby preventing cellular division. They are used in the treatment of psoriasis and cutaneous T-cell lymphoma among other skin and blood diseases. We have investigated the cytotoxic potential of three psoralen derivatives plus UVA exposure (PUVA) on a established cell line of human melanoma. Cells were treated with different concentrations of 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP) and 7-methylpyridopsoralen (MPP), for 1 h and after exposure to UVA light (0.3 J/cm(2)) were allowed to recover over a 24-72 h period. Viability was assessed by the microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Cisplatin, one of the most important drugs in the chemotherapy of melanoma, was included for comparative studies. All the psoralen derivatives tested were markedly cytotoxic in a dose and post-exposure-time dependent manner. The IC(50) values for 72 h of post-exposure time were as follows: MPP=0.05+/-0.01, TMP=0.13+/-0.003 and 8-MOP=10.79+/-1.85 micromol/L. Regardless of the limitations of the in vitro model, our results suggested that the lower IC(50) values of TMP and MPP might be of clinical importance.
Assuntos
Furocumarinas/farmacologia , Melanócitos/efeitos dos fármacos , Melanócitos/efeitos da radiação , Raios Ultravioleta , Divisão Celular/efeitos dos fármacos , Divisão Celular/efeitos da radiação , Linhagem Celular , Cisplatino/farmacologia , Colorimetria/métodos , Reagentes de Ligações Cruzadas , DNA/efeitos dos fármacos , DNA/efeitos da radiação , Relação Dose-Resposta a Droga , Humanos , Melanócitos/metabolismo , Metoxaleno/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Neoplasias Cutâneas/patologia , Sais de Tetrazólio/análise , Células Tumorais CultivadasRESUMO
A method for sample preparation and analysis by high performance liquid chromatography with UV detection (HPLC-UV) has been developed for routine analysis of psoralen and bergapten, photosensitizing compounds, in oral solutions of phytomedicines employed in Brazil for some illnesses. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. Calibration curves for psoralen and bergapten were linear in the range of 1.0-600.0 microg ml(-1) and 1.0-400.0 microg ml(-1) respectively. The recoveries of the psoralens in the oral solutions analysed were 94.43-99.97%. The percentage coefficient of variation (CV) of the quantitative analysis of the psoralens in the products analysis was within 5%. In inter-equipment study was employed gas chromatography-flame ionization (CG-FID) detection.
Assuntos
Furocumarinas/análise , Metoxaleno/análogos & derivados , Fármacos Fotossensibilizantes/análise , Plantas Medicinais/química , 5-Metoxipsoraleno , Calibragem , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Ionização de Chama , Metoxaleno/análise , Soluções Farmacêuticas , Reprodutibilidade dos Testes , Espectrofotometria UltravioletaRESUMO
8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 micro M). The intracellular Ca2+ chelator BAPTA/AM (1 micro M) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 micro M protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92% increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.
Assuntos
Melanoma/metabolismo , Metoxaleno/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Canais de Potássio Cálcio-Ativados/efeitos dos fármacos , Proteínas Tirosina Quinases/efeitos dos fármacos , Humanos , Indóis/farmacologia , Melanoma/patologia , Canais de Potássio Cálcio-Ativados/fisiologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/fisiologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Espectrometria de Fluorescência , Fatores de Tempo , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 µM). The intracellular Ca2+ chelator BAPTA/AM (1 µM) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 µM protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92 percent increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.
Assuntos
Humanos , Melanoma , Metoxaleno , Fármacos Fotossensibilizantes , Canais de Potássio , Proteínas Tirosina Quinases , Indóis , Transdução de Sinais , Espectrometria de Fluorescência , Fatores de Tempo , Células Tumorais CultivadasRESUMO
The use of furanocoumarins, which are photosensitizing compounds, combined with exposure to UV-A radiation is a common treatment for vitiligo, psoriasis, and a number of other skin diseases. Although furanocoumarins plus UV-A treatment is highly effective, several studies have shown that exposure to high doses increases the risk to development of cutaneus carcinoma. Several Dorstenia species are used in folk medicine, mainly against skin diseases, because of the presence of biologically active compounds. We present here analysis of the chemical composition of furanocoumarins from infusion and decoction of "Carapiá" (Dorstenia species), which is used in Brazil against several diseases. We have employed high-performance liquid chromatography (HPLC) procedures for the quantitative determination of psoralen, bergapten, and isopimpinellin. The contents of furanocoumarins revealed an insignificant difference between infusion and decoction. Dorstenia tubicina and D. asaroides contained psoralen and bergapten only in the rhizomes, whereas D. vitifolia shows solely isopimpinellin in both rhizomes and aerial parts.