Mestrado em Bioquímica
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- ItemAção da mitoquinona, um antioxidante mitocondrial, sobre o metabolismo mitocondrial cardíaco após infarto agudo do miocárdio(Universidade Federal do Espírito Santo, 2025-02-26) Traichel, Geórgia Azevedo Oliveira; Stefanon, Ivanita ; https://orcid.org/0000-0003-2638-5183; http://lattes.cnpq.br/8456612999765726; Fernandes, Aurélia Araújo; https://orcid.org/0000-0001-9945-1909; http://lattes.cnpq.br/5478728158150003; https://orcid.org/0009-0002-9695-3414; http://lattes.cnpq.br/0106389990223853; Vieira, Maicon Landim; https://orcid.org/0000-0002-6696-8888; http://lattes.cnpq.br/5216391153452438; Amorim, Girlandia Alexandre Brasil ; https://orcid.org/0000-0002-5455-7141; http://lattes.cnpq.br/1402295792093274Ischemic heart disease is one of the leading causes of heart failure (HF), with mitochondria playing a central role in the progression of this condition. Mitochondrial oxidative stress, alterations in cardiac bioenergetic pathways, and the activation of apoptotic signaling in cardiac tissue are key contributors of this process. In this context, identifying therapeutic strategies capable of directly targeting mitochondrial dysfunction is essential. This study aimed to evaluate the effects of mitoquinone (MitoQ) on mitochondrial function in infarcted rats treated for seven days. Male Wistar rats (aged 10 to 12 weeks) were randomly assigned to three groups: Sham, MI and MI MitoQ. Animals underwent surgery for myocardial infarction (MI) induction or a sham surgical procedure. Mitoquinone (MitoQ) treatment was administered in drinking water for seven days. Cardiac function was assessed by echocardiography, and weight data were analysed. As expected, the MI group showed a reduced ejection fraction and right ventricular hypertrophy, effects that were not reversed by the treatment. However, MitoQ attenuated pulmonary congestion. Additionally, the MI group exhibited an approximately 50% reduction in mitochondrial protein yield (YIELD), which was not restored with short-term treatment. Nevertheless, MitoQ improved mitochondrial calcium retention, enhanced mitochondrial coupling and ATP synthesis capacity, optimized complex I function, and reestablished beta-oxidation. Therefore, our findings suggest that MitoQ was effective in mitigating changes associated with mitochondrial dysfunction and may represent a promising complementary therapeutic strategy for ischemic heart disease, potentially supporting cardiac remodeling after infarction
- ItemPotencial efeito neuroprotetor do canabidiol em um modelo pré-clínico da doença de parkinson(Universidade Federal do Espírito Santo, 2024-10-18) Toledo, João Gabriel Assis; Pires, Rita Gomes Wanderley; https://orcid.org/0000-0002-4739-8349; https://orcid.org/http://lattes.cnpq.br/8356031036198869; https://orcid.org/0009-0008-9696-8679; http://lattes.cnpq.br/1148485070900089; Hollais, André Willian; https://orcid.org/0000-0002-2991-8646; http://lattes.cnpq.br/0068789063590867; Silva, Sarah Martins Presti da; https://orcid.org/0000-0003-2579-4866; http://lattes.cnpq.br/0768873502799973Parkinson's disease (PD) is the second most common neurodegenerative disease, second only to Alzheimer's disease. PD is characterized by the death of dopaminergic neurons in the substantia nigra pars compacta, leading to a decrease in dopamine release in the striatum region. The increase in cases of death and disability from PD has increased more than any other neurodegenerative disease, however, there is still no cure or treatment that reduces the progression of PD, making the search for new treatment methods and drug candidates that promote such effects imperative. The gold standard medication for the treatment of Parkinson's disease, levodopa, brings with it some problems with prolonged use, such as levodopa-induced dyskinesia, which is often worse than the disease itself. In this context, Cannabidiol (CBD), a non-psychoactive compound extracted from the Cannabis sativa plant, has shown promise, as there are several studies demonstrating its potential neuroprotective effect in different neurological diseases and also a possible indication for the treatment of levodopa-induced dyskinesia. Swiss mice were treated for 21 days with rotenone and two hours later cannabidiol was administered, and at the end of the treatment motor tests (open field and rotarod) were performed for biochemical analyses of the content of dopamine, serotonin and their metabolites by HPLC, in addition to the expression of tyrosine hydroxylase by Western Blotting. From the results obtained, there was no statistical difference in the results found in the open field test, however there was a significant difference in the latency to fall from the rotarod test apparatus, however, this difference was not found with the biochemical parameters both by Western Blotting and by HPLC. The data found in this work suggest a behavioral alteration that was not reflected in biochemical alterations and new experiments should be carried out with other doses of both rotenone and CBD.
- ItemPurificação e caracterização bioquímica de uma protease do muco do peixe escorpião scorpaena plumieri(Universidade Federal do Espírito Santo, 2024-10-09) Pirovani, Milenna Machado; Gonçalves, Juliana Barbosa Coitinho; Figueiredo, Suely Gomes de; https://orcid.org/0000-0002-5363-8329; Paula, Heberth de; Menezes, Thiago Nunes deProteases are the efficient executioners of a common chemical reaction, the hydrolysis of peptide bonds. These enzymes appear in all domains of life, including plants, animals, and microorganisms. Due to their involvement in various important biological processes (e.g., blood coagulation, blood pressure regulation, protein processing, and antimicrobial activity), as well as their high specificity, these enzymes have clinical and industrial biotechnological potential. Recently, using a proteomic approach – ‘shotgun’ modality – our research group identified 228 proteins in the epidermal mucus of the Brazilian scorpionfish, Scorpaena plumieri, including various classes of proteases. In this study, a gelatinolytic protease from S. plumieri skin mucus (named Sp-MGP) was purified to homogeneity through a combination of two chromatographic steps: conventional gel filtration (Sephacryl S-200 column) and anion exchange HPLC (SynChropak AX300 column). The purification process was monitored by zymography (SDS-PAGE gelatin). Physicochemical studies indicated that the purified enzyme is an approximately 79-kDa monomeric glycoprotein, as estimated by SDS-PAGE under both reducing and non-reducing conditions. Zymogram analysis of Sp-MGP showed gelatinolytic zones of activity in the gel, with optimal activity at pH 6.0 to 9.0, which is typical of serine proteases. The enzyme stored in aqueous solutions at room temperature (25–30°C), 4°C, -25°C, and -80°C for 60 days did not lose its activity. Sp-MGP was also active on the thrombin (S-2238) and kallikrein (S-2266) synthetic substrates. The activity on S2238 was inhibited by aprotinin and benzamidine. These data reaffirm the suggestion that the purified enzyme is a serine protease, specifically of the thrombin-like and kallikrein-like subclasses. The information generated represents the initial step in the study of a novel protease and may contribute to exploring its biotechnological potential, in addition to providing knowledge about the diversity of molecules in fish skin mucus.
- ItemEstudo do potencial antitumoral do extrato hidrolisado da espécie euphorbia tirucalli l. em modelos in vitro em câncer de ovário(Universidade Federal do Espírito Santo, 2024-07-30) Wanzeler, Ranna Batista; Rangel, Leticia Batista Azevedo; https://orcid.org/0000-0002-7810-0621; Tavares, Marcella Porto; Soares, Karla LirioCancer is a group of diseases characterized by the disordered proliferation of cells, which can be influenced by various factors. Ovarian cancer (OC) is the second most common gynecological neoplasm and the fifth most common cause of cancer-related death in women, primarily due to the asymptomatic growth of tumors. Cancer treatment involves various approaches, such as surgery, chemotherapy, and radiotherapy. However, chemoresistance is a challenge in the treatment of malignant neoplasms, leading to increased research on therapeutic combinations that include natural compounds. One substance that has been gaining attention is euphol, a compound found in the latex of the Euphorbia tirucalli plant. There are reports of its use in folk medicine to treat conditions such as inflammation, asthma, cough, warts, and malignant tumors. Thus, this study aimed to evaluate the antitumor potential of the hydrolyzed extract of Euphorbia tirucalli in vitro models using OC cell lines (A2780, ACRP, and OVCAR-3). Metabolic cell viability (MCV) experiments and IC₅₀ calculations were performed for OVCAR-3, A2780, and ACRP cell lines. Clonogenic assay, cell migration, and cell cycle analysis were performed only with OVCAR-3 cell line. MCV assay results showed that the A2780 and ACRP cell lines were not sensitive to the Euphorbia hydrolyzed extract, while the MCV of OVCAR-3 cell line had a significant decrease. Subsequent assays demonstrated that the hydrolyzed extract interferes with both migration and colony formation, drastically reducing viability. Finally, the cell cycle assay performed by flow cytometry showed that the cells were arrested in the G1 phase, corroborating other studies. Therefore, these results suggest a potential antitumor effect of the hydrolyzed extract of Euphorbia tirucalli.
- ItemAvaliação Físico-Química Da Interação De Betaciclodextrina Com Isoformas Alfa- E Psi- Tripsina(Universidade Federal do Espírito Santo, 2024-02-26) Oliveira, Daniel de Jesus de; Rosa, Dayanne Pinho; https://orcid.org/0000-0002-1483-5848; http://lattes.cnpq.br/7576001371663181; Santos, Alexandre Martins Costa; https://orcid.org/0000-0002-8801-8875; http://lattes.cnpq.br/4144105396879016; https://orcid.org/0009-0007-0082-9547; http://lattes.cnpq.br/5786173815850998; Araujo, Marlonni Maurastoni ; https://orcid.org/0000-0002-6064-3126; http://lattes.cnpq.br/6052184072658200; Cicilini, Maria Aparecida ; https://orcid.org/0000-0003-2751-117X; http://lattes.cnpq.br/7082425279468970Cyclodextrins are cyclic polysaccharides with a structure composed of glycopyranose. Studies have shown that cyclodextrins can form promising interactions with proteins, which has several applications in biotechnology and biochemistry, such as the solubilization of hydrophobic compounds and the protection of compounds sensitive to thermal, oxidative degradation, among others. In this work, in order to evaluate the influence of this polysaccharide on conformational stability and protein activity, physicochemical tools were used, adopting bovine trypsin as an enzymatic model of study, due to the knowledge of its structure and enzymatic activity. This enzyme belongs to the class of serine proteases and has several isoforms, α-trypsin, β-trypsin and ψ-trypsin being the most studied. The results showed that protease amidasic activity increased up to about 76% in commercial trypsin, 82% in alpha isoform, and 45% in psi isoform, suggesting that the addition of β-cyclodextrin optimized the amidsis activity of the trypsin isoforms tested. Fluorescence and absorption spectroscopy revealed changes in the structural properties of trypsin due to the interaction with βcyclodextrin, affecting the activity and conformational stability of trypsin. The formation of supramolecular states of trypsin isoforms in the presence of β-cyclodextrin by means of dynamic light scattering (DLS) was also evaluated, in which an increase in the size and dispersivity of the molecule suggestive of the formation of supramolecular states was verified. The joint evaluation of these results suggested that there was the formation of stabilizing interactions between β-cyclodextrin and trypsin isoforms through the formation of supramolecular states, with possible formation of inclusion complexes. Thus, it is suggested that β-cyclodextrin may interact with trypsin and affect its proteolytic activity and stability and thus the influence of the osmolyte can be understood at the molecular level.