RESUMO
The adult zebrafish is considered a useful model for studying mechanisms involved in tissue growth and regeneration. We have characterized cytotoxic damage to the retina of adult zebrafish caused by the injection of cobalt chloride (CoCl2 ) into the vitreous cavity. The CoCl2 concentration we used primarily caused injury to photoreceptors. We observed the complete disappearance of cones, followed by rods, across the retina surface from 28 to 96 hr after CoCl2 injury. The loss of 30% of bipolar cells was also observed by 50 hr after lesion (hpl). CoCl2 injury provoked a strong induction of the proliferative activity of multipotent Müller glia and derived progenitors. The effect of CoCl2 on retina cells was significantly reduced by treatment with glutamate ionotropic receptor antagonists. Cone photoreceptor regeneration occurred 25 days after injury. Moreover, a single dose of CoCl2 induced vascular damage and regeneration, whereas three injections of CoCl2 administered weekly provoked neovascular-like changes 20 days after injury. CoCl2 injury also caused microglial reactivity in the optic disc, retina periphery and fibre layer. CoCl2 -induced damage enhanced pluripotency and proneural transcription factor gene expression in the mature retina 72 hpl. Tumour necrosis factor alpha, vascular endothelial growth factor (VEGF) and VEGF receptor mRNA levels were also significantly enhanced by 72 hpl. The injury paradigm we have described in this work may be useful for the discovery of signalling molecules and pathways that participate in the regenerative response and it may serve as a model to screen for compounds that could potentially treat aberrant angiogenesis.
Assuntos
Proliferação de Células/fisiologia , Cobalto/toxicidade , Neovascularização Fisiológica/fisiologia , Neuroglia/metabolismo , Células Fotorreceptoras/metabolismo , Retina/metabolismo , Fatores Etários , Animais , Antimutagênicos/toxicidade , Proliferação de Células/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Regeneração Nervosa/fisiologia , Neuroglia/efeitos dos fármacos , Neuroglia/patologia , Células Fotorreceptoras/efeitos dos fármacos , Células Fotorreceptoras/patologia , Retina/efeitos dos fármacos , Retina/patologia , Peixe-ZebraRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Copaifera malmei Harms (Fabaceae), known mainly as óleo-mirim, is a native and endemic plant found in the states of Mato Grosso and Goiás of Brazil. The plant's leaves infusion is popularly used by riverine communities of the northern Araguaia microregion, Mato Grosso, Brazil, for the treatment of gastric ulcers and inflammatory diseases of the respiratory tract. The gastric antiulcer activity of the standardized leaves infusion extract of Copaifera malmei (SIECm) in rodents has been reported. The objective of this study was to advance the investigation of the safety profile of SIECm by evaluating the genotoxicity and subchronic toxicity using in vitro and in vivo experimental models. MATERIALS AND METHODS: SIECm was prepared by infusion, by incubating the powdered dried leaves material in boiled water for 15min. In vitro genotoxicity of SIECm (10, 30 or 100µg/mL) was assessed by micronucleus and comet tests using Chinese hamster ovary (CHO-k1) epithelial cells. The evaluation of subchronic toxicity profile was performed by daily oral administration of SIECm (100, 400 or 1000mg/kg) to Wistar rats for 30 days. Clinical observations of toxicological related parameters were done every 6 days. After the treatment period, blood was collected for hematological and biochemical analysis, and some organs were removed for macroscopic and histopathological analysis. RESULTS: In the micronucleus assay, SIECm demonstrated anti-mutagenic activity. In the comet assay, SIECm presented anti-genotoxic effect preventing DNA damage at all the three concentrations tested with pre-treatment, while the same effect was only observed in the co-treatment at the lowest concentration. Post-treatment with SIECm increased the genetic damage induced by hydrogen peroxide (H2O2) at the highest concentration. In the subchronic toxicity test, few changes were observed, such as increase in feed consumption in the group of animals treated with 100mg/kg of the SIECm, which reversed after 6 days. There were no macroscopic, histological and relative weights changes in the organs of animals treated with SIECm. No toxicologically relevant changes were observed in the hematological analysis. Subchronic administration of SIECm reduced levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in animals treated with 100mg/kg and serum triglyceride levels at 400 and 1000mg/kg. However, the hematological and biochemical changes observed are within the physiological ranges for this animal species. CONCLUSION: The results demonstrate that SIECm is not genotoxic, and does not present toxicity when used orally for up to 30 days. In addition, it showed protection to the genetic damage induced by H2O2. The SIECm therefore has a high safety margin for therapeutic use.
Assuntos
Antimutagênicos/toxicidade , Fabaceae , Extratos Vegetais/toxicidade , Animais , Células CHO , Ensaio Cometa , Cricetulus , Dano ao DNA/efeitos dos fármacos , Feminino , Peróxido de Hidrogênio/toxicidade , Testes para Micronúcleos , Folhas de Planta , Ratos Wistar , Testes de Toxicidade SubcrônicaRESUMO
Gentisic acid (GA) exhibits antioxidant, anti-inflammatory, and antibiotic activities. This substance can be found in citrus fruits, grapes, olive oil, and peas. Considering that there are few studies in the literature on the toxicity of GA, the present work aimed to investigate its cytotoxic, mutagenic, and antimutagenic activities on HTC cells. GA was diluted in culture medium at the final concentration of 0.08, 0.16, 0.8, 1.6, and 8 µg/mL. The cytotoxicity was determined by the MTT assay and Trypan Blue exclusion method, with methyl methanesulfonate and doxorubicin as positive controls, respectively. The cytokinesis-block micronucleus assay determined the mutagenic/antimutagenic activity with benzo[a]pyrene as positive control. Negative control received culture medium only. GA (0.08-8 µg/mL) was not cytotoxic to HTC cells by the MTT assay nor the Trypan Blue exclusion method as no statistical difference was observed when compared to the control. Concentration of 0.08 and 0.8 µg/mL showed no mutagenic or clastogenic effects, as no significant micronuclei inductions were observed, different from 8 µg/mL, that was mutagenic. Furthermore, none of the concentrations presented an antiproliferative activity. The antimutagenic activity of GA (0.08 µg/mL) was observed at the simultaneous treatment, as it reduced the frequency of micronuclei by 76% (24 h) and 79% (48 h). Although pre- and post-treatments were not statistically different from the mutagen, they reduced the induced-damage by 11% and 21%, respectively. The present study indicated the absence of cytotoxicity and antiproliferative activities of GA, in addition to their antimutagenic/protective effects that may contribute to human health.
Assuntos
Antimutagênicos/farmacologia , Gentisatos/farmacologia , Hepatócitos/efeitos dos fármacos , Mutagênicos/farmacologia , Animais , Antimutagênicos/toxicidade , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Gentisatos/toxicidade , Hepatócitos/metabolismo , Hepatócitos/patologia , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Testes para Micronúcleos , Mutagênicos/toxicidade , Ratos , Medição de Risco , Fatores de TempoRESUMO
CONTEXT: Helicteres vegae Cristóbal (Sterculiaceae) (Hv) and Heliopsis sinaloensis B.L. Turner (Asteraceae) (Hs) are endangered and poorly studied plant species; related plants have been used against chronic-degenerative and infectious diseases. Therefore, Hv and Hs could be sources of bioactive compounds against these illnesses. OBJECTIVE: To determine the chemical composition and biological activities (antioxidant, antimutagenic and antimicrobial) of Hv and Hs leaves (L) and stems (S). MATERIALS AND METHODS: Methanol extracts (ME) of each plant/tissue were evaluated for their phytochemicals; phenolics (HPLC-DAD-ESI-MS); antioxidant activity (AA) (0.125-4 mg/mL) (DPPH, ABTS, ORAC and ß-carotene discoloration); antimutagenicity (0.5 and 1 mg/plate) (Ames assay, tester strain Salmonella enterica serovar Typhimurium YG1024, 1-nitropyrene as mutagen); activity against human pathogens (1 mg/mL); and toxicity (0.01-2 mg/mL) (Artemia salina assay). RESULTS: All ME showed flavonoids and triterpenes/steroids. The ME-SHv had the highest content of total phenolics (TP) (2245.82 ± 21.45 mg GAE/100 g d.w.) and condensed tannins (603.71 ± 1.115 mg CE/100 g d.w.). The compounds identified were flavonoids (kaempferol 7-O-coumaroylhexoside, and two kaempferol 7-O-rhamnosylhexosides) and phenolics [rosmarinic acid, and 3'-O-(8â³-Z-caffeoyl) rosmarinic acid]. The ME-LHs showed the highest content of flavonoids (357.88 mg RE/g d.w.) and phenolic acids (238.58 mg CAE/g d.w.) by HPLC. The ME-SHv showed the highest AA. All ME were strong antimutagens (63.3-85.7%). Only the Hs extracts were toxic (ME-LHs, LC50 = 94.9 ± 1.7 µg/mL; ME-SHs, LC50 = 89.03 ± 4.42 µg/mL). DISCUSSION AND CONCLUSIONS: Both Hv and Hs are potential sources of preventive and therapeutic agents against chronic-degenerative diseases.
Assuntos
Anti-Infecciosos/farmacologia , Antimutagênicos/farmacologia , Antioxidantes/farmacologia , Asteraceae/química , Malvaceae/química , Extratos Vegetais/farmacologia , Animais , Anti-Infecciosos/química , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/toxicidade , Antimutagênicos/química , Antimutagênicos/isolamento & purificação , Antimutagênicos/toxicidade , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/toxicidade , Artemia/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Benzotiazóis/química , Compostos de Bifenilo/química , Cromatografia Líquida de Alta Pressão , Giardia lamblia/efeitos dos fármacos , Giardia lamblia/crescimento & desenvolvimento , Metanol/química , Testes de Sensibilidade Microbiana , Testes de Mutagenicidade , Capacidade de Absorbância de Radicais de Oxigênio , Testes de Sensibilidade Parasitária , Fitoterapia , Picratos/química , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Folhas de Planta/química , Caules de Planta/química , Plantas Medicinais , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Solventes/química , Espectrometria de Massas por Ionização por Electrospray , Ácidos Sulfônicos/química , beta Caroteno/químicaRESUMO
Machaerium hirtum (Vell.) Stellfeld (M.hirtum) is a plant known as 'jacarandá-bico-de-pato' whose bark is commonly used against diarrhea, cough and cancer. The aim of this study was to phytochemically characterise the hydroethanolic extract of this plant, investigate its antimutagenic activities using the Ames test and evaluate its effects on cell viability, genomic instability, gene expression and cell protection in human hepatocellular carcinoma cells (HepG2). Antimutagenic activity was assessed by simultaneous pre- and post-treatment with direct and indirect mutagens, such as 4-nitro-o-phenylenediamine (NPD), mitomycin C (MMC), benzo[a]pyrene (B[a]P) and aflatoxin B1 (AFB1), using the Ames test, cytokinesis blocking micronucleus and apoptosis assays. Only 3 of the 10 concentrations evaluated in the MTT assay were cytotoxic in HepG2 cells. Micronucleated or apoptotic cells were not observed with any of the tested concentrations, and there were no mutagenic effects in the bacterial system. However, the Nuclear Division Index and flow cytometry data showed a decrease in cell proliferation. The extract showed an inhibitory effect against direct (NPD) and indirect mutagens (B[a]P and AFB1). Furthermore, pre- and post-treated cells showed significant reduction in the number of apoptotic and micronucleated cells. This effect is not likely to be associated with the modulation of antioxidant genes, as shown by the RT-qPCR results. Six known flavonoids were identified in the hydroethanolic extract of Machaerium hirtum leaves, and their structures were elucidated by spectroscopic and spectrophotometric methods. The presence of the antioxidants apigenin and luteolin may explain these protective effects, because these components can inhibit the formation of reactive species and prevent apoptosis and DNA damage. In conclusion, the M.hirtum extract showed chemopreventive potential and was not hazardous at the tested concentrations in the experiments presented here. Moreover, this extract should be investigated further as a chemopreventive agent.
Assuntos
Antimutagênicos/farmacologia , Fabaceae/química , Extratos Vegetais/farmacologia , Antimutagênicos/química , Antimutagênicos/toxicidade , Apoptose/efeitos dos fármacos , Apoptose/genética , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Linhagem Celular Tumoral , Dano ao DNA/efeitos dos fármacos , Flavonoides/química , Flavonoides/farmacologia , Flavonoides/toxicidade , Citometria de Fluxo , Expressão Gênica , Humanos , Micronúcleos com Defeito Cromossômico , Testes para Micronúcleos , Testes de Mutagenicidade , Mutação/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/toxicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genéticaRESUMO
The search for substances able to inhibit and/or diminish the effects of genotoxic and mutagenic substances has been the target of several investigations performed in recent times. Hymenoptera venoms constitute a considerable source of substances with pharmacological potential. The present study aimed to evaluate the cytotoxic, genotoxic and anti-genotoxic, mutagenic and anti-mutagenic potentials of Apis mellifera venom in HepG2 cells. In this evaluation, the MTT test was applied to determine the most appropriate concentrations for the genotoxicity and mutagenicity tests. It was verified that the concentrations of 0.1, 0.05 and 0.01µg/mL were not cytotoxic, hence these concentrations were used in the experiments. For the evaluation of the genotoxic and mutagenic potential of the bee venom the comet assay and the micronucleus test were applied, respectively. The concentrations mentioned above presented both genotoxic and mutagenic potential for HepG2 cells and it was necessary to test lower concentrations of the venom (10pg/mL, 1pg/mL and 0.1pg/mL) for the anti-genotoxicity and anti-mutagenicity tests, which were performed subjecting the cells to the action of MMS (methyl methanesulfonate) in order to verify the ability of the venom to inhibit or diminish the action of this compound, which has a recognized action on the genetic material. Pre-, post-treatment and simultaneous treatment with and without incubation with the venom were performed. It was observed that the lowest three concentrations tested did not present any anti-genotoxic and anti-mutagenic activity on the cells. The use of bee venom for pharmacological purposes in treatments such as cancer must be done with extreme caution, since it was observed that even at very low concentrations the venom can induce genotoxicity and mutagenicity in human cells, as was verified for the HepG2 cells.
Assuntos
Antimutagênicos/farmacologia , Venenos de Abelha/farmacologia , Dano ao DNA , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Animais , Antimutagênicos/toxicidade , Venenos de Abelha/toxicidade , Abelhas , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Metanossulfonato de Metila/análogos & derivados , Metanossulfonato de Metila/toxicidade , Testes de Mutagenicidade , Mutagênicos/toxicidadeRESUMO
The possible benefits of some bioactive flavones and xanthones present in plants of the genus Syngonanthus prompted us to screen them for estrogenic activity. However, scientific research has shown that such substances may have undesirable properties, such as mutagenicity, carcinogenicity and toxicity, which restrict their use as therapeutic agents. Hence, the aim of this study was to assess the estrogenicity and mutagenic and antimutagenic properties. We used recombinant yeast assay (RYA), with the strain BY4741 of Saccharomyces cerevisiae, and Ames test, with strains TA100, TA98, TA97a and TA102 of Salmonella typhimirium, to evaluate estrogenicity, mutagenicity and antimutagenicity of methanolic extracts of Syngonanthus dealbatus (S.d.), Syngonanthus macrolepsis (S.m.), Syngonanthus nitens (S.n.) and Syngonanthus suberosus (S.s.), and of 9 compounds isolated from them (1=luteolin, 2=mix of A-1,3,6-trihydroxy-2-methoxyxanthone and B-1,3,6-trihydroxy-2,5-dimethoxyxanthone, 3=1,5,7-trihydroxy-3,6-dimethoxyxanthone, 4=1,3,6,8-tetrahydroxy-2,5-dimethoxyxanthone, 5=1,3,6,8-tetrahydroxy-5-methoxyxanthone, 6=7-methoxyluteolin-8-C-ß-glucopyranoside, 7=7-methoxyluteolin-6-C-ß-glucopyranoside, 8=7,3'-dimethoxyluteolin-6-C-ß-glucopyranoside and 9=6-hydroxyluteolin). The results indicated the estrogenic potential of the S. nitens methanol extract and four of its isolated xanthones, which exhibited, respectively, 14.74±1.63 nM; 19.54±6.61; 7.20±0.37; 6.71±1.02 e 10.01±4.26 nM of estradiol-equivalents (EEQ). None of the extracts or isolated compounds showed mutagenicity in any of the test strains and all of them showed antimutagenic potential, in particular preventing mutations caused by aflatoxin B1 (AFB1) and benzo[a]pyrene (B[a]P). The results show that the xanthones, only isolated from the methanol extract of S. nitens capitula, probably were the responsible for its estrogenic activity and could be useful as phytoestrogens, providing a new opportunity to develop hormonal agents. In addition, flavones and xanthones could also be used as a new antimutagenic agent. Since, the mutagens are involved in the initiation and promotion of several human diseases, including cancer, the significance of novel bioactive phytocompounds in counteracting these pro-mutagenic and carcinogenic effects is now gaining credence.
Assuntos
Antimutagênicos/farmacologia , Eriocaulaceae/química , Estrogênios/farmacologia , Flavonas/farmacologia , Xantonas/farmacologia , Antimutagênicos/isolamento & purificação , Antimutagênicos/toxicidade , Quimioprevenção , Estrogênios/isolamento & purificação , Estrogênios/toxicidade , Flavonas/isolamento & purificação , Flavonas/toxicidade , Humanos , Metanol/química , Mutagênicos/toxicidade , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Saccharomyces cerevisiae/efeitos dos fármacos , Salmonella/efeitos dos fármacos , Xantonas/isolamento & purificação , Xantonas/toxicidadeRESUMO
BACKGROUND: St. John's wort (Hypericum perforatum L.) is an herbaceous plant that is native to Europe, West Asia and North Africa and that is recognized and used worldwide for the treatment of mild and moderate depression. It also has been shown to be therapeutic for the treatment of burns, bruises and swelling and can be used for its wound healing, antiviral, antimicrobial, antioxidant, analgesic, hepato-protective and anxiolytic properties. The aim of this study was to evaluate the potential cytotoxic, mutagenic and antimutagenic action of H. Perforatum. METHODS: Meristematic cells were used as the test system for Allium cepa L., and bone marrow cells from Rattus norvegicus, ex vivo, were used to calculate the mitotic index and the percentage of chromosomal aberration. Statistical analysis was performed using the chi-square test. RESULTS: This medicinal plant had no cytotoxic potential in the vegetal test system evaluated. In the animal test system, none of the acute treatments, including intraperitoneal gavage and subchronic gavage, were cytotoxic or mutagenic. Moreover, this plant presented antimutagenic activity against the clastogenic action of cyclophosphamide, as confirmed in pre-treatment (76% reduction in damage), simultaneous treatment (95%) and post-treatment (97%). CONCLUSIONS: Thus, the results of this study suggest that the administration of H. perforatum, especially by gavage similar to oral consumption used by humans, is safe and with beneficial antimutagenic potential.
Assuntos
Antidepressivos/toxicidade , Antimutagênicos/toxicidade , Hypericum/química , Mutagênicos/toxicidade , Cebolas/efeitos dos fármacos , Extratos Vegetais/toxicidade , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Aberrações Cromossômicas/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Masculino , Mitose/efeitos dos fármacos , Cebolas/citologia , Ratos , Ratos WistarRESUMO
Diphenyl diselenide (DPDS) is an electrophilic reagent used in the synthesis of a variety of pharmacologically active organoselenium compounds. Studies have shown its interesting pharmacodinamic properties, as antioxidant, antimutagenic and antitumoral effects. Here we report the antigenotoxic properties of DPDS against tamoxifen (TAM)-induced oxidative DNA damage in MCF-7 cultured cell line. We determined the cytotoxicity by lactate dehydrogenase (LDH) leakage assay and evaluated oxidative DNA damage by modified comet assay employing the enzymes formamidopyrimidine DNA-glycosylase (Fpg) and endonuclease III (Endo III). Our results demonstrate that the cellular effects of DPDS appear to be complex and concentration-dependent. The present findings show that DPDS is not genotoxic (at concentrations lower than 2.0µmol/L) in MCF-7 cells, as observed in the modified comet assay. Moreover, DPDS protects against TAM-induced oxidative DNA damage, probably by its antioxidant activity, without interfering with its cytotoxicity. In this manner, the treatment with low concentrations of DPDS, a synthetic organoselenium compound, could be used as a potent antigenotoxic agent to prevent the risk of cancer induction triggered by tamoxifen hormone therapy. Thereby, more studies concerning the toxicity of DPDS and its structural derivatives are still necessary for future safe therapeutic application and development of novel chemopreventive compounds for combined therapy in breast cancer.
Assuntos
Antimutagênicos/farmacologia , Antineoplásicos Hormonais/toxicidade , Derivados de Benzeno/farmacologia , Compostos Organosselênicos/farmacologia , Tamoxifeno/toxicidade , Antimutagênicos/administração & dosagem , Antimutagênicos/toxicidade , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Antioxidantes/toxicidade , Derivados de Benzeno/administração & dosagem , Derivados de Benzeno/toxicidade , Neoplasias da Mama/patologia , Ensaio Cometa , Dano ao DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Células MCF-7 , Compostos Organosselênicos/administração & dosagem , Compostos Organosselênicos/toxicidade , Estresse Oxidativo/efeitos dos fármacosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Brassica oleracea L. var. acephala D.C. has been extensively used in Brazilian traditional medicine to treat gastric ulcer. AIM OF THE STUDY: This study was conducted to evaluate the in vivo genotoxic and/or antigenotoxic potential of a Brassica oleraceae hydroalcoholic extract obtained from the leaves, in different cells of mice. MATERIALS AND METHODS: Analyses were performed using the comet assay, on leukocytes (collected 4 and 24 h after treatment), liver, brain, bone marrow and testicular cells (collected 24 h after treatment), and using the micronucleus test (MN) in bone marrow cells. Eight groups of albino Swiss mice were treated (N=6): control (C), positive control (doxorubicin 80 mg/kg (DXR)), and six experimental groups, which received 500, 1000 and 2000 mg/kg of Brassica oleraceae extract alone by gavage, while a further three groups received the same doses plus DXR (80 mg/kg). We calculated the damage scores, and their averages were compared by ANOVA followed by the Tukey test for multiple comparisons. RESULTS: The results demonstrated that none of the tested doses of Brassica oleraceae extract showed genotoxic effects by the comet assay, or clastogenic effects by the MN test. On the other hand, for all cells evaluated, the three tested doses of the Brassica extract promoted inhibition of DNA damage induced by DXR. CONCLUSIONS: Under our experimental conditions, Brassica oleraceae leaf extract showed no genotoxic or clastogenic effects in different cells of mice. However, it did show a significant decrease in DNA damage induced by doxorubicin. It is suggested that the antigenotoxic properties of this extract may be of great pharmacological importance, and may be beneficial for cancer prevention.
Assuntos
Antimutagênicos/toxicidade , Brassica , Extratos Vegetais/toxicidade , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Células Cultivadas , Ensaio Cometa , DNA/efeitos dos fármacos , Doxorrubicina/toxicidade , Masculino , Camundongos , Testes para Micronúcleos , Folhas de PlantaRESUMO
Myrciaria dubia, a plant native to the Amazon region, stands out as a fruit rich in vitamin C and other metabolites with nutritional potential. We evaluated the antioxidant, genotoxic and antigenotoxic potential of M. dubia juice on blood cells of mice after acute, subacute and chronic treatments. Flavonoids and vitamin C present in the fruit of M. dubia were quantified. In vitro antioxidant activity was evaluated by DPPH assay. Blood samples were collected for analysis after treatment, and the alkaline comet assay was used to analyze the genotoxic and antigenotoxic activity (ex vivo analysis using H(2)O(2)). The amount of vitamin C per 100mL of M. dubia was 52.5mg. DPPH assay showed an antioxidant potential of the fruit. No M. dubia concentration tested exerted any genotoxic effect on mice blood cells. In the ex vivo test, the juice demonstrated antigenotoxic effect, and acute treatment produced the most significant results. After the treatments, there was no evidence of toxicity or death. In conclusion, our data show that M. dubia juice has antigenotoxic and antioxidant activities, though with no genotoxicity for blood cells. Nevertheless, more in-depth studies should be conducted to assess the safety of this fruit for human consumption.
Assuntos
Antimutagênicos/farmacologia , Células Sanguíneas/efeitos dos fármacos , Myrtaceae/química , Extratos Vegetais/farmacologia , Animais , Antimutagênicos/toxicidade , Ensaio Cometa , Feminino , Masculino , Camundongos , Mutagênicos/toxicidade , Testes de Toxicidade Aguda , Testes de Toxicidade Crônica , Testes de Toxicidade SubagudaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Fabiana species (Solanaceae family) extracts have long been used in Argentinean traditional medicine as anti-inflammatories, antiseptic, bone fractures and others diseases, but there is no scientific evidence which supports their use. AIM OF THE STUDY: The present study was conducted to evaluate the ability of aqueous and ethanolic extracts of four Fabiana species (Fabiana bryoides Phil., Fabiana punensis A.C. Arroyo, Fabiana densa J. Rèmy and Fabiana patagonica Speg.) to inhibit key enzymes in inflammatory processes, free radical scavenging properties and genotoxic effects. MATERIALS AND METHODS: HPLC-DAD of aqueous and ethanolic extracts from four Fabiana species was established. All Fabiana extracts were evaluated on their ability to inhibit hyaluronidase and lipoxygenase enzymes to assess their activity against inflammatory mediators. Antioxidant capacity was determined using the 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assays and ß-carotene-linolenic acid assay. Genotoxicity was evaluated by the Ames assay. RESULTS: The results indicated that the chromatographic patterns of four Fabiana species were different in quantity and absorption intensity of peaks. The alcoholic extract of Fabiana punensis was the most active scavenger of DPPH and ABTS(+) radicals (SC(50) values of 3.85 ± 0.24 and 2.56 ± 0.10 µgGAE/mL, respectively). Fabiana patagonica extracts exhibited the highest peroxyl radical scavenging activity compared with the other three taxa (IC(50) values between 1.00 ± 0.04 and 4.46 ± 0.40 µg GAE/mL for all extracts) and anti-lipoxygenase activity with IC(50) values between 12.5 and 15.5 µg GAE/mL. The absence of mutagenicity indicates that the DNA does not seem to be a relevant target for these extracts. Fabiana bryoides ethanolic extract showed an interesting effect: it inhibited spontaneous mutagenesis, which could be considered as an antimutagenic effect in the TA98 (+S9) and TA100 (+S9/-S9) strains. The potency differences found between the species could be consequence of the different phytochemical pattern observed by HPLC. CONCLUSIONS: The inhibitory effects on lipoxygenase and hyaluronidase, free radical scavenging activities and lack of genotoxicity of Fabiana extracts may support the folk use of Fabiana punensis, Fabiana patagonica, Fabiana bryoides and Fabiana densa as inhibitor of inflammatory mediators.
Assuntos
Anti-Inflamatórios/farmacologia , Antimutagênicos/farmacologia , Etanol/química , Sequestradores de Radicais Livres/farmacologia , Extratos Vegetais/farmacologia , Solanaceae , Solventes/química , Água/química , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/toxicidade , Antimutagênicos/química , Antimutagênicos/isolamento & purificação , Antimutagênicos/toxicidade , Argentina , Benzotiazóis , Compostos de Bifenilo/química , Cromatografia Líquida de Alta Pressão , Dano ao DNA , DNA Bacteriano/efeitos dos fármacos , Relação Dose-Resposta a Droga , Etnofarmacologia , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/toxicidade , Hialuronoglucosaminidase/antagonistas & inibidores , Hialuronoglucosaminidase/metabolismo , Cinética , Inibidores de Lipoxigenase/farmacologia , Medicina Tradicional , Picratos/química , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Plantas Medicinais , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Solanaceae/química , Ácidos Sulfônicos/química , Tiazóis/química , Ácido alfa-Linolênico/química , beta Caroteno/químicaRESUMO
Solanum lycocarpum A. St. Hill. (Family Solanaceae), popularly known in Brazil as lobeira, is a common weed in the Brazilian Cerrado vegetation. The fruits of this species have been used in Brazil for culinary purposes and in folk medicine as a sedative, diuretic, antiepileptic, antispasmodic, hypoglycemic, and hypocholesterolemic agent as well as in the control of obesity. Due to the spreading use of this plant as a therapeutic resource and food, the present study aimed to evaluate the genotoxic, antigenotoxic, and cytotoxic effects of S. lycocarpum ethanolic fruit extract using the mouse bone marrow micronucleus test. Both genotoxicity and antigenotoxicity of this ethanolic fruit extract were evaluated by using the frequency of micronucleated polychromatic erythrocytes, whereas cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio. Our results indicated that although S. lycocarpum ethanolic fruit extract did not exhibit genotoxic effect in mice bone marrow, both cytotoxic and antigenotoxic actions were evidenced at all tested doses.
Assuntos
Antimutagênicos/farmacologia , Citotoxinas/toxicidade , Frutas/química , Mutagênicos/toxicidade , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Solanum/química , Algoritmos , Animais , Animais não Endogâmicos , Antimutagênicos/toxicidade , Células da Medula Óssea/efeitos dos fármacos , Brasil , Citotoxinas/farmacologia , Relação Dose-Resposta a Droga , Masculino , Medicina Tradicional/efeitos adversos , Camundongos , Testes para Micronúcleos , Mitomicina/antagonistas & inibidores , Mitomicina/toxicidade , Mutagênicos/farmacologia , Distribuição Aleatória , Testes de Toxicidade AgudaRESUMO
Biflorin is a natural quinone isolated from Capraria biflora L. Previous studies demonstrated that biflorin inhibits in vitro and in vivo tumor cell growth and presents potent antioxidant activity. In this paper, we report concentration-dependent cytotoxic, genotoxic, antimutagenic, and protective effects of biflorin on Salmonella tiphymurium, yeast Saccharomyces cerevisiae, and V79 mammalian cells, using different approaches. In the Salmonella/microsome assay, biflorin was not mutagenic to TA97a TA98, TA100, and TA102 strains. However, biflorin was able to induce cytotoxicity in haploid S. cerevisiae cells in stationary and exponential phase growth. In diploid yeast cells, biflorin did not induce significant mutagenic and recombinogenic effects at the employed concentration range. In addition, the pre-treatment with biflorin prevented the mutagenic and recombinogenic events induced by hydrogen peroxide (H(2)O(2)) in S. cerevisiae. In V79 mammalian cells, biflorin was cytotoxic at higher concentrations. Moreover, at low concentrations biflorin pre-treatment protected against H(2)O(2)-induced oxidative damage by reducing lipid peroxidation and DNA damage as evaluated by normal and modified comet assay using DNA glycosylases. Our results suggest that biflorin cellular effects are concentration dependent. At lower concentrations, biflorin has significant antioxidant and protective effects against the cytotoxicity, genotoxicity, mutagenicity, and intracellular lipid peroxidation induced by H(2)O(2) in yeast and mammalian cells, which can be attributed to its hydroxyl radical-scavenging property. However, at higher concentrations, biflorin is cytotoxic and genotoxic.
Assuntos
Antimutagênicos/toxicidade , Antineoplásicos/toxicidade , Gleiquênias/química , Naftoquinonas/toxicidade , Animais , Antimutagênicos/química , Antimutagênicos/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Linhagem Celular , Ensaio Cometa , Ensaios de Seleção de Medicamentos Antitumorais , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/toxicidade , Naftoquinonas/química , Naftoquinonas/isolamento & purificação , Saccharomyces cerevisiae/efeitos dos fármacos , Salmonella/efeitos dos fármacosRESUMO
Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.
Assuntos
Antimutagênicos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Fígado/efeitos dos fármacos , Mutagênicos/toxicidade , Extratos Vegetais/farmacologia , Pueraria/química , Salmonella typhimurium/efeitos dos fármacos , Animais , Antimutagênicos/isolamento & purificação , Antimutagênicos/toxicidade , Bacillus subtilis/genética , Masculino , Testes para Micronúcleos/métodos , Mutagênicos/isolamento & purificação , Extratos Vegetais/toxicidade , Ratos , Ratos Sprague-Dawley , Salmonella typhimurium/genética , Espectrofotometria , Fatores de TempoRESUMO
Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.
Assuntos
Animais , Masculino , Ratos , Antimutagênicos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Fígado/efeitos dos fármacos , Mutagênicos/toxicidade , Extratos Vegetais/farmacologia , Pueraria/química , Salmonella typhimurium/efeitos dos fármacos , Antimutagênicos/isolamento & purificação , Antimutagênicos/toxicidade , Bacillus subtilis/genética , Testes para Micronúcleos/métodos , Mutagênicos/isolamento & purificação , Extratos Vegetais/toxicidade , Ratos Sprague-Dawley , Espectrofotometria , Salmonella typhimurium/genética , Fatores de TempoRESUMO
Ischemic brain injury is a dynamic process that involves oxidative stress, inflammation, and cell death, as well as activation of endogenous adaptive and regenerative mechanisms depending on activation of transcription factors such as hypoxia inducible factor 1-alpha (HIF-1alpha). Because CoCl2 activates HIF-1alpha, we described a new focal-hypoxia model by direct intracerebral CoCl2 injection. Adult male Wistar rats were intracerebrally injected with CoCl2 (2 microl-50 mM), in frontoparietal cortex of right hemisphere, and saline (2 microl) in the contralateral hemisphere. In slides of fixed brains at 1, 6, 9, 24 h or 5 day after treatment, TTC, histochemistry (toluidine blue, Hoescht-33342, TUNEL), immunostaining (HIF-1alpha, GFAP), Lycopersicon esculentum lectin staining, and electron microscopy (EM) were performed. Immediately after 1 h post CoCl2 injection, HIF-1alpha stabilization and neuronal nuclear shrinkage and cromathin condensation were observed by immunostaining and EM, respectively. Neuronal apoptotic nuclear morphology and GFAP immunoreactivity and lectin maximal reactivity were detected during 6-9 h. Ultrastructural alterations of morphology included edematous perinuclear cytoplasm, organelles and endoplasmic reticulum (RE) enlargement, mitochondrial swelling with increased matrix density, and deposits of electron-dense material. Neurons showed particular nuclear indentations. Astrocytes and oligodendrocytes presented alterations in both nuclei and RE with dilated lumen and altered mitochondrias, and all these ultrastructural changes became detectable at day 5. CoCl2 cortical injection mimics focal brain ischemia, inducing neuronal death and glial activation. This model brings the opportunity to develop focal ischemia in selected brain areas to study their functional consequences and potential pharmacological therapies for in vivo models of stroke.
Assuntos
Antimutagênicos/toxicidade , Cobalto/toxicidade , Hipóxia/induzido quimicamente , Hipóxia/patologia , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Análise de Variância , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Morte Celular/efeitos dos fármacos , Modelos Animais de Doenças , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Marcação In Situ das Extremidades Cortadas/métodos , Lectinas/efeitos dos fármacos , Masculino , Microscopia Eletrônica de Transmissão/métodos , Proteínas do Tecido Nervoso/metabolismo , Neuroglia/ultraestrutura , Neurônios/ultraestrutura , Ratos , Ratos Wistar , Coloração e Rotulagem/métodos , Fatores de TempoRESUMO
The marine environment is a rich source of biological active compounds and the sponges can be considered the most productive one. This diversity gives rise to unique chemical compounds with potential pharmacological properties. Our study is focused on the genotoxic and antigenotoxic evaluation of two crude extracts obtained from the Brazilian endemic marine sponge Arenosclera brasiliensis. Salmonella typhimurium reverse mutation test with TA97, TA98, TA100 and TA102 strains were performed. For antimutagenic analysis, a pre-, co-, and post-treatment to evaluate, respectively, intracellular and extracellular reactions and possible modulation on DNA repair. Additionally, in order to verify the influence of the crude extracts on DNA damage induction, a plasmid-DNA treatment was assayed. No mutagenicity was observed in Salmonella reverse mutation test, neither DNA strand induced damage. Antimutagenic activity was observed in pre-, co-, and post-treatment. A significant antigenotoxic effect was observed in the crude extract, which suggests that A. brasiliensis extract has the potential to protect DNA from the action of 4NQO, 2-aminofluorene, sodium azide and mitomycin C.
Assuntos
Antimutagênicos/toxicidade , Poríferos/química , Salmonella typhimurium/efeitos dos fármacos , Animais , Antimutagênicos/isolamento & purificação , Brasil , Dano ao DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Mutagênicos/isolamento & purificação , Mutagênicos/toxicidade , Plasmídeos/metabolismo , Salmonella typhimurium/genética , Extratos de TecidosRESUMO
Senna (Cassia angustifolia Vahl.) is widely used as a laxative, although potential side effects, such as toxicity and genotoxicity, have been reported. This study evaluated genotoxic and mutagenic effects of senna aqueous extract (SAE) by means of four experimental assays: inactivation of Escherichia coli cultures; bacterial growth inhibition; reverse mutation test (Mutoxitest) and DNA strand break analysis in plasmid DNA. Our results demonstrated that SAE produces single and double strand breaks in plasmid DNA in a cell free system. On the other hand, SAE was not cytotoxic or mutagenic to Escherichia coli strains tested. In effect, SAE was able to avoid H(2)O(2)-induced mutagenesis and toxicity in Escherichia coli IC203 (uvrA oxyR) and IC205 (uvrA mutM) strains, pointing to a new antioxidant/antimutagenic action of SAE.
Assuntos
Mutagênicos/toxicidade , Extrato de Senna/toxicidade , Antimutagênicos/farmacologia , Antimutagênicos/toxicidade , Antioxidantes/farmacologia , Antioxidantes/toxicidade , Quebras de DNA de Cadeia Dupla/efeitos dos fármacos , Quebras de DNA de Cadeia Simples/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Peróxido de Hidrogênio/metabolismo , Técnicas In Vitro , Testes de Mutagenicidade/métodos , Mutagênicos/farmacologia , Plasmídeos/efeitos dos fármacos , Plasmídeos/metabolismo , Extrato de Senna/farmacologia , Senna/químicaRESUMO
Vanillin (VA), the world's major flavoring compound used in food industry and confectionery products - that has antimutagenic and anticarcinogenic activity against a variety of mutagenic/carcinogenic agents - was tested for the interval between the formation of premutational lesion and it is finalization as a DNA lesion. The overall findings using co-treatment protocols in SMART test suggest that VA can lead to a significant protection against the general genotoxicity of ethylmethanesulphonate (EMS), N-ethyl-N-nitrosourea (ENU), N-methyl-N-nitrosourea (MNU) and bleomycin sulphate (BLEO). Considering MNU, ENU and EMS the desmutagenic activity observed could result from VA-stimulation of detoxification, via induction of glutathione S-transferase. However, the protector effect related to BLEO could be attributed to its powerful scavenger ability, which has the potential to prevent oxidative damage induced by BLEO.