RESUMO
BACKGROUND: Cuban sugarcane wax acids (SCWA) and policosanol (PCO) are mixtures of higher aliphatic acids and alcohols, respectively, purified from sugarcane wax with different chief components. Although it has been known that they have antioxidant and anti-inflammatory activities, physiological properties on molecular mechanism of SCWA have been less studied than PCO. METHODS: In this study, we compared antiatherogenic activities of SCWA and PCO via encapsulation with reconstituted high-density lipoproteins (rHDL). RESULTS: After reconstitution, SCWA-rHDL showed smaller particle size than PCO-rHDL with increase of content. PCO-rHDL or SCWA-rHDL showed distinct inhibition of glycation with similar extent in the presence of fructose. PCO-rHDL or SCWA-rHDL showed strong antioxidant activity against cupric ion-mediated oxidation of low-density lipoproteins (LDL), and inhibition of oxLDL uptake into macrophages. Although PCO-rHDL showed 1.2-fold stronger inhibition against cholesteryl ester transfer protein (CETP) activity than SCWA-rHDL, SCWA-rHDL enhanced 15% more brain cell (BV-2) growth and 23% more regeneration of tail fin in zebrafish. CONCLUSION: PCO and SCWA both enhance the beneficial functions of HDL to maximize its antioxidant, antiglycation, and antiatherosclerotic activities and the inhibition of CETP. These enhancements of HDL functionality by PCO and SCWA could exert antiaging and rejuvenation activity.
Assuntos
Ácidos/farmacologia , Anticolesterolemiantes/farmacologia , Proteínas de Transferência de Ésteres de Colesterol/antagonistas & inibidores , Álcoois Graxos/farmacologia , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/metabolismo , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Saccharum/química , Ceras/química , Ácidos/isolamento & purificação , Nadadeiras de Animais/efeitos dos fármacos , Nadadeiras de Animais/crescimento & desenvolvimento , Animais , Anticolesterolemiantes/isolamento & purificação , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proteínas de Transferência de Ésteres de Colesterol/metabolismo , Álcoois Graxos/isolamento & purificação , Humanos , Macrófagos/metabolismo , Masculino , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Oxirredução , Extratos Vegetais/isolamento & purificação , Regeneração , Células THP-1 , Adulto Jovem , Peixe-Zebra/crescimento & desenvolvimentoRESUMO
The cytochrome P450 1A (CYP1A) mRNA is induced by environmental contaminants such as PAHs, PCBs and dioxins. The present study cloned the CYP1A transcript from the guppy Phalloceros caudimaculatus, which represents a potential fish for toxicological studies in South America. The newly identified CYP1A encodes a protein with 521 amino acids that shared 96-70% identity with other fishes. The characterization of organ- and time-dependent induction of CYP1A using RT-qPCR was evaluated after waterborne exposure to beta-naphthoflavone (BNF; 1µM). The minimum exposure time that elicited significant CYP1A induction was 1h for liver, gill, gut, brain, anal fin and fingerlings; 2h for dorsal fin; and 4h for kidney and tail fin. CYP1A tended to reach peak induction in the first few hours (4h-8h) of experiment in most organs, although levels remained induced until the end of the experiment (96h). Validation of CYP1A use in environmental sample was performed by exposing P. caudimaculatus to elutriate made from sediment of three streams located in adjacent areas of the Patos Lagoon Estuary (RS, Brazil). CYP1A in liver, gills and anal fin was induced by elutriate made from urban (S1) and industrial (S2) sites; and not induced by a reference site located 22 Km from potential contaminant sources, suggesting that environmental contamination plays a role in this induction. The results suggest that fins could be used for CYP1A biomarker analysis and employed in non-lethal biopsy methods for environmental monitoring. The responsiveness of the newly identified CYP1A to BNF and elutriate indicates that the guppy P. caudimaculatus could be used for environmental toxicology investigations in South American environments.
Assuntos
Citocromo P-450 CYP1A1/metabolismo , Poecilia/metabolismo , RNA Mensageiro/metabolismo , Poluentes Químicos da Água/toxicidade , beta-Naftoflavona/toxicidade , Sequência de Aminoácidos , Nadadeiras de Animais/efeitos dos fármacos , Nadadeiras de Animais/metabolismo , Animais , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/química , Citocromo P-450 CYP1A1/genética , Monitoramento Ambiental , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Dados de Sequência Molecular , Poecilia/crescimento & desenvolvimento , Alinhamento de Sequência , Poluentes Químicos da Água/químicaRESUMO
The level of cytochrome P450 1A (CYP1A) in fish is used as a typical environmental biomarker for the presence of organic contaminants. We used RT-qPCR to investigate CYP1A mRNA levels in the liver, gill and gonopodium of guppies Jenynsia multidentata and Phalloceros caudimaculatus in wetlands within the Rio Grande city (RG) which is under the influence of the Patos Lagoon Estuary (RS, Brazil). The CYP1A mRNA levels evaluated in fish liver from two locations that receive non-treated wastewater effluents (S3 and S4) and another locations near an oil refinery (S6) and an industrial complex (S7), were higher than in locations remote from those sites (S1, S2 and S5). The sum of 16 priority PAHs in sediment confirmed high levels in S4 and S6 (3914.0 and 4414.0 ng g(-1) dw, respectively) comparing to S7>S2>S3>S5>S1 (119.3, 66.3, 62.8, 16.4 and 1.7 ng g(-1) dw). J. multidentata from sites S1 to S4 that were transferred to the laboratory exhibited CYP1A induction after 24 h waterborne exposure to 1 µM betanaphtoflavone (BNF) in all organs compared to controls, except in the liver of fish from site S4. This lack of CYP1A induction by BNF indicates a CYP1A refractory phenotype in guppy. Although this characteristic possibly involves the alteration in AHR signaling or control, the mechanism of resistance is unknown. The present study provides information about the use of the use of CYP1A in South American guppies as an useful biomarker tool for environmental contamination studies.
Assuntos
Ciprinodontiformes/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Monitoramento Ambiental/métodos , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análise , Nadadeiras de Animais/efeitos dos fármacos , Nadadeiras de Animais/enzimologia , Animais , Biomarcadores/metabolismo , Brasil , Ciprinodontiformes/genética , Citocromo P-450 CYP1A1/genética , Expressão Gênica/efeitos dos fármacos , Sedimentos Geológicos/química , Brânquias/efeitos dos fármacos , Brânquias/enzimologia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Fenótipo , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Água do Mar/química , Poluentes Químicos da Água/toxicidadeRESUMO
Studies have demonstrated that the prolonged use of corticoids can delay the healing process, affecting re-epithelialization, neovascularization and collagen synthesis. As the fins of teleost fish contain a large amount of collagen, the aim of the present study was to investigate the effect of dexamethasone (anti-inflammatory and glucocorticoid steroid widely used in the treatment of rheumatic diseases) during the regeneration process in the caudal fin of specimens of carp (Cyprinus carpio). For such, two glass aquaria were used - one for a group of fish treated with dexamethasone (Henrifarma) in a 20 mg/L concentration and the other for the control group. The caudal fins were amputated transversally and fish remained in their respective aquaria until regeneration occurred. Samples of regenerating fins were collected on days 1, 2, 4, 6, 8 and 10 after amputation. The fins in the control group regenerated normally and grew within the expected in time course. The fins in the group treated with dexamethasone were significantly smaller in comparison to the control group at every evaluation time. Thus, it was possible to verify that, at this concentration of dexamethasone, the regeneration of the caudal fins was delayed, but not completely inhibited. The results show that the caudal fin is a good model for histological studies on regeneration and the action of drug toxicity, but it's also of great importance the interaction with further studies for a better knowledge and understanding of all the changes in all the phases.
Assuntos
Nadadeiras de Animais/efeitos dos fármacos , Carpas/fisiologia , Dexametasona/farmacologia , Regeneração/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Nadadeiras de Animais/fisiologia , Animais , Estudos de Casos e Controles , Fatores de Tempo , Cicatrização/fisiologiaRESUMO
Studies have demonstrated that the prolonged use of corticoids can delay the healing process, affecting re-epithelialization, neovascularization and collagen synthesis. As the fins of teleost fish contain a large amount of collagen, the aim of the present study was to investigate the effect of dexamethasone (anti-inflammatory and glucocorticoid steroid widely used in the treatment of rheumatic diseases) during the regeneration process in the caudal fin of specimens of carp (Cyprinus carpio). For such, two glass aquaria were used – one for a group of fish treated with dexamethasone (Henrifarma) in a 20 mg/L concentration and the other for the control group. The caudal fins were amputated transversally and fish remained in their respective aquaria until regeneration occurred. Samples of regenerating fins were collected on days 1, 2, 4, 6, 8 and 10 after amputation. The fins in the control group regenerated normally and grew within the expected in time course. The fins in the group treated with dexamethasone were significantly smaller in comparison to the control group at every evaluation time. Thus, it was possible to verify that, at this concentration of dexamethasone, the regeneration of the caudal fins was delayed, but not completely inhibited. The results show that the caudal fin is a good model for histological studies on regeneration and the action of drug toxicity, but it’s also of great importance the interaction with further studies for a better knowledge and understanding of all the changes in all the phases.
Estudos mostram que corticóides usados por longos períodos podem atrasar o processo de cicatrização, influenciando na reepitelização, na neovascularização e na síntese do colágeno. Os constituintes das nadadeiras dos peixes teleósteos contêm grande quantidade de colágeno e assim o objetivo do presente trabalho foi estudar o efeito da dexametasona (um antiinflamatório e glicocorticóide esteróide bastante utilizado no tratamento de doenças reumáticas) durante o processo regenerativo das nadadeiras caudais das carpas (Cyprinus carpio). Para isso, foram montados dois aquários de vidro, um para o grupo controle e outro para o grupo tratado com a dexametasona (Henrifarma) na concentração de 20mg/L. Os peixes distribuídos nesses aquários tiveram suas nadadeiras caudais amputadas transversalmente e permaneceram nos respectivos aquários para que ocorresse a regeneração. Foram feitas coletas das nadadeiras em regeneração em intervalos de 1, 2, 4, 6, 8 e 10 dias após a amputação. Foi observado que nos peixes do grupo controle, as nadadeiras regeneraram normalmente e cresceram o esperado em cada intervalo de tempo. No entanto, foi verificado que nos peixes do grupo tratado com dexametasona, em cada intervalo analisado, as nadadeiras regeneradas dos peixes expostos à droga eram menores que a medida das nadadeiras dos peixes do grupo controle. Assim, foi possível verificar que, nessa concentração de dexametasona, a regeneração das nadadeiras caudais foi mais lenta, mas não ocorreu a total inibição da regeneração. Dessa forma, os resultados comprovam que a nadadeira caudal é um bom modelo para estudos histológicos sobre a regeneração e a ação da toxicidade de drogas, mas, também, é de grande importância a interação com estudos mais aprofundados para se conhecer e compreender melhor todas as alterações em todas as fases.
Assuntos
Animais , Nadadeiras de Animais/efeitos dos fármacos , Carpas/fisiologia , Dexametasona/farmacologia , Regeneração/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Nadadeiras de Animais/fisiologia , Estudos de Casos e Controles , Fatores de Tempo , Cicatrização/fisiologiaRESUMO
Cytochrome P450 1A (CYP1A) expression in fish is used as a biomarker of exposure to organic contaminants, such PAHs, PCBs and dioxins, in the aquatic environment. South American guppy fish Jenynsia multidentata were exposed to the prototypical aryl hydrocarbon receptor (AHR) agonist beta-naphthoflavone (BNF; 1µM) and the fins were biopsied to characterize different aspects of CYP1A induction. RTq-PCR was used to quantify CYP1A mRNA levels in fish tissues. CYP1A induction in the gill, liver and anal fin (gonopodium) occurred within the first hour of waterborne exposure to BNF and persisted throughout 2, 4, 8, 24, 48 and 96h compared to controls (DMSO vehicle; p<0.05). The organ-specific temporal pattern of induction was marked by mRNA levels consistently augment as duration of exposure increases and tend to a sustained induction from 24h to 96h for gill and liver (â¼15-fold and â¼50-fold over control, respectively). In gonopodium, there was a maximum CYP1A mRNA level at 4h (â¼34-fold over control). Basal CYP1A mRNA levels and its induction following BNF exposure were not affected by administration of a chemical anesthetic (fish immersion in 100mgl(-1) MS-222 for 2-5min) in the gill, liver, gonopodium, dorsal or tail fin (p<0.05). In an ex vivo assay, in which small pieces of biopsied fins were exposed to BNF for 4h, high CYP1A induction was observed in the tail and gonopodium (â¼49-fold and â¼69-fold, respectively) but not in the dorsal fin compared to controls. To our knowledge, this is the first study to show that a 1h waterborne exposure to an AHR agonist is sufficient to cause CYP1A induction in fish organs and fins. The present study added new information to the field regarding the use of MS-222 as an anesthetic on fish and the analysis of biopsied fins as an alternative non-lethalex vivo assay for evaluating the CYP1A biomarker in fish. This observation could be useful for planning fish toxicological bioassays and biomonitoring studies on the aquatic environments in South America.
Assuntos
Aminobenzoatos/farmacologia , Anestésicos/farmacologia , Citocromo P-450 CYP1A1/biossíntese , Poluentes da Água/toxicidade , beta-Naftoflavona/toxicidade , Nadadeiras de Animais/efeitos dos fármacos , Nadadeiras de Animais/enzimologia , Animais , Ciprinodontiformes/genética , Ciprinodontiformes/metabolismo , Citocromo P-450 CYP1A1/genética , Brânquias/efeitos dos fármacos , Brânquias/enzimologia , Cinética , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , RNA Mensageiro/biossíntese , Receptores de Hidrocarboneto Arílico/agonistasRESUMO
This study focused on several wetlands in Laguna del Tigre National Park (Guatemala) as part of Conservation International's Rapid Assessment Program. Sediment and water samples were collected from a laguna near Xan field, Guatemala's largest oil facility, and three other sites for determination of levels of polycyclic aromatic hydrocarbons (PAHs). Cichlid fish (Thorichthys meeki and Vieja synspila) were collected for determination of DNA strand breakage (by gel electrophoresis), chromosomal breakage (flow cytometry), and fin erosion. For T. meeki from Xan field, chromosomal breakage and strand breakage was greater than in at least two of the three reference sites. For V. synspila, chromosomal breakage and strand breakage were greater in Xan than one of the two reference sites. Fin erosion was observed only at the Xan laguna. Genetic biomarker effects and fin erosion, along with patterns of aqueous PAH concentrations, indicate that fish are affected by anthropogenic contaminants. PAHs were elevated at some reference sites, but environmental forensic analysis suggested a pyrogenic or diagenic origin. It is possible that oil field brines injected into the ground water caused fin erosion and genotoxicity in fish at Xan field, and it is also possible that pyrogenic PAHs influence levels of DNA damage in reference sites. These analyses represent one of the first efforts to examine genotoxicity in native Mesoamerican cichlids.