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Item Avaliação da atividade genotóxica e antigenotóxica de Celtis iguanaea (Jacq.) Sargent em bactérias e camundongos(Universidade Federal de Goiás, 2010-02-24) Borges, Flávio Fernandes Veloso; Lee, Chen Chen; http://lattes.cnpq.br/4621907105842007Celtis iguanaea (Jacq.) Sargent is a member of the Cannabaceae family and popularly known as esporão de galo. Ethnobotanical surveys of cerrado native plants show that the tea leaves of C. iguanaea is used in traditional medicine for body aches, asthma, cramps, poor digestion, urinary infections, kidney disfunctions, as a stimulant or as a diuretic.The purpose of the present work was to evaluate the possible cytotoxic, genotoxic and antigenotoxic effects of the leaves aqueous extract of C. iguanaea by the prophage λ induction test (SOS Inductest) and the micronucleous test in mice bone marrow. The SOS Inductest was performed according to Moreau (1976), using lysogenic WP2s(λ) and indicator RJF013 strains derived from Escherichia coli. WP2s(λ) cultures were treated with different doses of the extract (0.5, 1, 2.5, and 5 mg) (evaluation of genotoxicity) or co-treated with a single dose of MMC (0,5 mg) and different doses of extract (assessment of antigenotoxicity). Then, the treated culture was added to the indicator strain (RJF013) and both were poured on plates in LB(1/2) medium. For the evaluation of cytotoxicity, cultures of WP2s(λ) treated with different doses of extract were diluted in M9 buffer and plated on LB. The micronucleus test was carried out according to Schmid (1975). To assess the genotoxic activity, animals were treated with Celtis iguanaea extract (100, 300 and 500 mg/kg concentrations). To evaluate the antigenotoxic activity, the same doses of extract were treated simultaneously with 4mg/Kg of MMC. The frequencies of micronucleated polychromatic erythrocytes (MNPCE) were evaluated at 24h and 48h exposure period except the negative control (24h). The cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). In both tests the obtained results demonstrated anticytotoxic activity and absence of genotoxicity and cytotoxicity of the Celtis iguanaea leaves extract. The extract also showed partial antigenotoxic activity in bacteria and antigenotoxic activity in mice.Item Avaliação dos efeitos cardiovasculares do aceturato de diminazeno (DIZE) em ratos submetidos à sobrecarga pressórica(Universidade Federal de Goiás, 2014-02-12) Macedo, Larissa Matuda; Colugnati, Diego Basile; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314Activation of the Angiotensin Converting Enzyme 2 (ACE2)-Angiotensin-(1-7) [Ang(1-7)]-Mas Receptor axis results in protective effects in the cardiovascular system. ACE 2 is an important component of Renin-Angiotensin System, because it is able to convert Angiotensin II in Ang-(1-7). Recents studies have shown that diminazene aceturate (DIZE) act as an activator of ACE 2. The objective of this study was to evaluate the cardiovascular effects of chronic treatment with DIZE in pressureoverloaded rats and the possible mechanisms involved in these effects. Male Wistar rats (200-350 g) were divided in four groups: (1) Sham; (2) Coarcted (abdominal aortic banding, CAA); (3) CAA + DIZE (1 mg/kg, gavage); e (4) CAA + DIZE (1 mg/kg, gavage) + A-779 (120 µg/day, osmotic mini-pumps). Twenty one days after surgery procedure, the blood pressure was recorded, the hearts were isolated and perfused according to Langendorff method. Vascular reactivity was measured by isolated aortic ring preparation. In order to evaluate the cardiac hypertrophy, the left ventricular mass index (VMI) was calculated through the ratio of the left ventricular wet weight to tibia length. The cross-sectional area of cardiomyocytes (CSA) was also evaluated. The mRNA levels for ANP, BNP e TGF-β were also evaluated by qRT-PCR. The expression of ACE-2 and ERK1/2, AKT, mTOR, GATA-4, catalase and SOD proteins involved in hypertrophic pathways was analyzed by Western Blot technique. The results are presented as means ± SEM. One-way ANOVA followed by the Newman-Keuls post-test was used to analyze the blood pressure, cardiac morphometric parameters, isolated heart, qRT-PCR and Western Blot experiments. Two-way ANOVA followed by the Bonferroni post-test was used for aortic rings preparation protocols. All statistical analyses were considered significant at P<0.05. Isolated hearts from coarcted rats presented a significant decrease in the left ventricular end systolic pressure (128.1 ± 9.0 vs. 79.1 ± 12.8 mmHg in CAA, P<0.05), left ventricular developed pressure (118.1 ± 8.9 vs. 69.0 ± 12.7 mmHg in CAA, P<0.05), +dP/dt (2295.0 ± 161.8 vs. 1406.0 ± 246.5 mmHg/s in CAA, P<0.05) and dP/dt (1787.0 ± 166.0 vs. 1066.0 ± 181.9 mmHg/s in CAA, P<0.05). The DIZE treatment attenuated all of these effects induced by CAA. Moreover, DIZE treatment increased the coronary flow in hypertrophic hearts (CAA: 11.6 ± 0.8 vs. CAA+DIZE: 15.8 ± 0.6 mL/min, P<0.05). This effect was blocked by A-779. Pressure–overloaded hearts showed a significant increase in VMI (0.17 ± 0.01 vs. 0.21 ± 0.01 g/cm in CAA, P<0.05) and CSA (8.98 ± 0.54 vs. 10.72 ± 0.27 µm in CAA, P<0.05). The chronic treatment with DIZE prevented the heart hypertrophy (10.72 ± 0.27 in CAA vs. 9.25 ± 0.23 µm in CAA+DIZE, P<0.05). Indeed, treatment with A-779 attenuated the antihypertrophic effect induced by DIZE treatment. The coarcted rats presented a increase in mRNA expression of ANP, BNP and TGF-β and the DIZE treatment reverted this effect. The pressure overload decreased the acetylcholine-induced relaxation in aortic rings from coarcted rats and treatment with DIZE was not able to improve this effect. The coarctation decreased the phosphorylation of the AKT, which was not changed by DIZE treatment. The expression of ACE 2, total and phosphorylated ERK1/2, total AKT, mTOR, SOD and catalase was not changed by coarctation or by ACE 2 activation. These results show that the chronic treatment with DIZE was efficient in preventing the left ventricular hypertrophy and cardiac dysfunction induced by pressure overload. These effects were independent of changes in the expression of ACE 2, ERK1/2, AKT, mTOR, SOD and catalase. Thus, DIZE has important therapeutic potential for cardiovascular diseases.Item Efeito anti-proliferativo de curcumina associada á nanopartículas magnéticas funcionalizadas com bicamada de ácido láurico sobre células de melanoma humano skmel 37(Universidade Federal de Goiás, 2011) Souza, Fernanda França de; Guillo, Lídia Andreu; http://lattes.cnpq.br/3401436781775091Curcumin has emerged as a great promise for cancer treatment and chemoprophylaxis with curcumin, but its low solubility in water is minimal systemic bioavailability. Attempts were made to improve its solubility in aqueous solutions by incorporating curcumin in liposomes or micelles, but these systems have low stability. In addition, magnetic nanoparticles have been used as promising drug delivery systems because they can be functionalized to become water soluble and biocompatible. Studies with curcumin incorporated into the magnetic nanoparticles are still scarce. This study aims to evaluate the antiproliferative effect of curcumin combined with magnetic nanoparticles functionalized bilayer of lauric acid in human melanoma cell line SKMEL 37. Human melanoma cells were treated at different concentrations and cytotoxicity was evaluated by MTT assay. The cell morphology was evaluated by light microscopy and fluorescence. Our studies have shown that curcumin has incorporated into nanoparticles cytotoxic effect at concentrations above 40,8 μg/ml or 111 μM. Since free curcumin caused significant death at concentrations above 11.5 μM. We also observed morphological changes typical of apoptosis such as chromatin condensation, nuclear fragmentation and bleb formation. These findings show us that both their cytotoxic activity, as both the magnetic properties and its solubility in water make this new formulation with a curcumin compound interest for therapeutic use.Item Desenvolvimento de sistemas lipídicos nanoestruturados contendo paclitaxel: estudos de permeação cutânea(Universidade Federal de Goiás, 2014-01-29) Tosta, Fabiana Vaz; LIma, Eliana Martins; Taveira, Stephânia Fleury; http://lattes.cnpq.br/0382450621383005; Taveira, Stephânia Fleury; Marreto, Ricardo Neves; Lopes, Flávio MarquesPaclitaxel (PTX) is a natural product extracted from the bark of the Pacific Yew and has numerous antitumor actions, including skin cancers. The topical treatment of skin and pre-cancerous lesions cancer is desired, since the systemic treatment has many side effects However, PTX to be incorporated into formulations suitable for it to penetrate the stratum corneum and skin tumors reached. Lipid nanoparticles have potential to increase drug retention in the stratum corneum, thus providing controlled release and great percutaneous absorption. Within this context, the aim of this study was to develop and characterize solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) containing the antineoplastic PTX and evaluate its permeation into the pig ear skin in vertical diffusion cells type "Franz". Quantification of PTX paclitaxel was performed by high performance liquid chromatography. The NLS and CLNs were obtained by the method of dilution of the microemulsion containingcetylpyridinium chloride, glyceryl behenate, triglycerides of caprylic / capric acid, polysorbate 80 and sorbitan trioleate 85. The particles were characterized by medium size, PdI, zeta potential, encapsulation efficiency, drug loading and recovery. Stability studies were carried out for a period of 30 days with storage at 4 °C (± 2 °C). Theskin permeation studies of the PTX nanoparticles were conducted in “Franz” type diffusion cells in pig ear skin. The NLS obtained showed average size of 314.1 ± 10.9 to 335.9 ± 0.9nm. The CLN obtained with more oil in the lipid matrix (CLN100)showed average size 270.6 ± 13.5 nm. The encapsulation efficiency of the systemsobtained was above 90% when 3.75% was added PTX formulations. The stability studies revealed a trend in increasing the size of the particles PdI along the storage period, but these differences are not statistically significant. The CLN100 increased about 3 times the amount of drug in the stratum corneum (SC) as compared to the administration of unencapsulated drug and also increased by 1.5 times the amount of PTX in the SC in relation to the topical application of other lipid particles. Thus, the lipid particles appear to be promising systems for topical application of PTX.