Navegando por Autor "Serquiz, Raphael Paschoal"

Agora exibindo 1 - 6 de 6

Biochemical characterisation of a kunitz-type inhibitor from tamarindus indica L. seeds and its efficacy in reducing plasma leptin in an experimental model of obesity
(Journal of Enzyme Inhibition and Medicinal Chemistry, 2018-01) Maciel, Bruna Leal Lima; Medeiros, Amanda Fernandes de; Costa, Izael de Sousa; Carvalho, Fabiana Maria Coimbra de; Kiyota, Sumika; Souza, Beatriz Blenda Pinheiro de; Sifuentes, Daniel Nogoceke; Serquiz, Raphael Paschoal; Uchôa, Adriana Ferreira; Santos, Elizeu Antunes dos; Morais, Ana Heloneida de Araújo
A trypsin inhibitor isolated from tamarind seed (TTI) has satietogenic effects in animals, increasing the cholecystokinin (CCK) in eutrophy and reducing leptin in obesity. We purified TTI (pTTI), characterised, and observed its effect upon CCK and leptin in obese Wistar rats. By HPLC, and after amplification of resolution, two protein fractions were observed: Fr1 and Fr2, with average mass of [M þ 14H]þ ¼ 19,594,690 Da and [M þ 13H]þ ¼ 19,578,266 Da, respectively. The protein fractions showed 54 and 53 amino acid residues with the same sequence. pTTI presented resistance to temperature and pH variations; IC50 was 2.7 1010mol.L1 and Ki was 2.9 1011mol.L1 . The 2-DE revealed spots with isoelectric points between pH 5 and 6, and one near pH 8. pTTI action on leptin decrease was confirmed. We conclude that pTTI is a Kunitz trypsin inhibitor with possible biotechnological health-related application
Inibidor de serinoproteinase obtido de sementes de Juquiri (Mimosa regnelli Benth.) com atividade anti-inflamatória, anticoagulante e adjuvante da heparina
(2017-10-25) Serquiz, Raphael Paschoal; Santos, Elizeu Antunes dos; ; ; Morais, Ana Heloneida de Araújo; ; Bezerra, Fabiana Lima; ; Dore, Celina Maria Pinto Guerra;
A hemostasia é o evento pelo qual os mamíferos bloqueiam a perda de volemias sanguíneas e promovem o reparo de lesões, e falhas por excessos contribuem para a propagação de fatores inflamatórios e trombóticos. As consequências da aterosclerose são exemplos de agravos desse tipo e a heparina (Hep) é um fármaco de escolha para esses tratamentos. Pacientes com carência de antitrombina têm resistência aos efeitos anticoagulantes da Hep, então pesquisas por novas moléculas homeostáticas que contribuam com os tratamentos clínicos são necessárias. Inibidores de proteinases vegetais têm sido eficazes em controlar processos biológicos, como coagulação e inflamação. O presente trabalho objetivou isolar um inibidor de tripsina da semente de Juquiri (JTI) verificando sua atividade contra serinoproteinases, e avaliar seus potenciais anticoagulante e anti-inflamatório. O inibidor foi isolado por cromatografia de afinidade e troca-iônica, apresentando duas principais bandas proteicas de 11,9 e 19,2 kDa estimadas por SDS-PAGE, e foi capaz de inibir tripsina e quimotripsina. Na coagulação, o JTI prolongou o tempo de tromboplastina parcial (APTT) em concentrações superiores à 2 µg/100 µL de plasma, sem prolongar o tempo de protrombina (PT). Associado à Hep, o JTI foi capaz de intensificar o efeito anticoagulante do fármaco em ambos os testes. Sem apresentar toxicidade contra eritrócitos humanos o JTI reduziu 40% das concentrações de elastase liberada por neutrófilos induzidos por PAF (fator ativador de plaquetas). O JTI em concentrações entre 20-80 µg/mL estimulou a produção de TNF por macrófagos RAW 264.7 em cultura sem provocar outros estímulos inflamatórios. Macrófagos ativados por LPS tiveram redução nas concentrações de NO, IL-6 e TNF quando tratados com JTI em concentrações entre 20-160 µg/mL. Estes dados apontam o JTI como promissor no tratamento de doenças trombóticas e inflamatórias, tendo sido eficaz como adjuvante da heparina e em promover a redução de estímulos inflamatórios de neutrófilos por vias provavelmente dependentes de MAPK, e em macrófagos por vias do fator de transcrição NF-κB.
A lactose-binding lectin from the marine sponge cinachyrella apion (Cal) induces cell death in uuman cervical adenocarcinoma cells
(Marine Drugs, 2012-03) Morais, Ana Heloneida de Araújo; Rabelo, Luciana; Monteiro, Norberto de Kássio Vieira; Serquiz, Raphael Paschoal; Santos, Paula; Oliveira, Ruth; Oliveira, Adeliana Silva de; Rocha, Hugo; Uchôa, Adriana Ferreira; Santos, Elizeu Antunes dos
Cancer represents a set of more than 100 diseases, including malignant tumors from different locations. Strategies inducing differentiation have had limited success in the treatment of established cancers. Marine sponges are a biological reservoir of bioactive molecules, especially lectins. Several animal and plant lectins were purified with antitumor activity, mitogenic, anti-inflammatory and antiviral, but there are few reports in the literature describing the mechanism of action of lectins purified from marine sponges to induce apoptosis in human tumor cells. In this work, a lectin purified from the marine sponge Cinachyrella apion (CaL) was evaluated with respect to its hemolytic, cytotoxic and antiproliferative properties, besides the ability to induce cell death in tumor cells. The antiproliferative activity of CaL was tested against HeLa, PC3 and 3T3 cell lines, with highest growth inhibition for HeLa, reducing cell growth at a dose dependent manner (0.5–10 µg/mL). Hemolytic activity and toxicity against peripheral blood cells were tested using the concentration of IC50 (10 µg/mL) for both trials and twice the IC50 for analysis in flow cytometry, indicating that CaL is not toxic to these cells. To assess the mechanism of cell death caused by CaL in HeLa cells, we performed flow cytometry and western blotting. Results showed that lectin probably induces cell death by apoptosis activation by pro-apoptotic protein Bax, promoting mitochondrial membrane permeabilization, cell cycle arrest in S phase and acting as both dependent and/or independent of caspases pathway. These results indicate the potential of CaL in studies of medicine for treating cancer
Satietogenic protein from tamarind seeds decreases food intake, leptin plasma and CCK-1r gene expression in obese wistar rats
(Obesity Facts, 2018) Maciel, Bruna Leal Lima; Costa, Izael De Souza; Medeiros, Amanda Fernandes de; Carvalho, Fabiana Maria Coimbra de; Lima, Vanessa Cristina Oliveira de; Serquiz, Raphael Paschoal; Serquiz, Alexandre Coelho; Silbiger, Vivian Nogueira; Bortolin, Raul Hernandes; Santos, Elizeu Antunes dos
Objective: This study evaluated the effect of a protein, the isolated Trypsin Inhibitor (TTI) from Tamarindus indica L. seed, as a CCK secretagogue and its action upon food intake and leptin in obese Wistar rats. Methods: Three groups of obese rats were fed 10 days one of the following diets: Standard diet (Labina®) + water; High Glycemic Index and Load (HGLI) diet + water or HGLI diet + TTI. Lean animals were fed the standard diet for the 10 days. Food intake, zoometric measurements, plasma CCK, plasma leptin, relative mRNA expression of intestinal CCK-related genes, and expression of the ob gene in subcutaneous adipose tissue were assessed. Results: TTI decreased food intake but did not increase plasma CCK in obese animals. On the other hand, TTI treatment decreased CCK-1R gene expression in obese animals compared with the obese group with no treatment (p = 0.027). Obese animals treated with TTI presented lower plasma leptin than the non-treated obese animals. Conclusion: We suggest that TTI by decreasing plasma leptin may improve CCK action, regardless of its increase in plasma from obese rats, since food intake was lowest
Supplementation with a new trypsin inhibitor from peanut is associated with reduced fasting glucose, weight control, and increased plasma CCK secretion in an animal model
(Journal of Enzyme Inhibition and Medicinal Chemistry, 2016-02) Maciel, Bruna Leal Lima; Serquiz, Alexandre Coelho; Machado, Richele Janaína de Araújo; Serquiz, Raphael Paschoal; Lima, Vanessa Cristina Oliveira; Carvalho, Fabiana Maria Coimbra de; Carneiro, Marcella Araújo do Amaral; Uchôa, Adriana Ferreira; Santos, Elizeu Antunes dos; Morais, Ana Heloneida de Araújo
Ingestion of peanuts may have a beneficial effect on weight control, possibly due to the satietogenic action of trypsin inhibitors. The aim of this study was to isolate a new trypsin inhibitor in a typical Brazilian peanut sweet (pac¸oca) and evaluate its effect in biochemical parameters, weight gain and food intake in male Wistar rats. The trypsin inhibitor in peanut pac¸oca (AHTI) was isolated. Experimental diets were prepared with AIN-93G supplemented with AHTI. Animals had their weight and food intake monitored. Animals were anesthetized, euthanized, and their bloods collected by cardiac puncture for dosage of cholecystokinin (CCK) and other biochemical parameters. Supplementation with AHTI significantly decreased fasting glucose, body weight gain, and food intake. These effects may be attributed to increased satiety, once supplemented animals showed no evidence of impaired nutritional status and also because AHTI increased CCK production. Thus, our results indicate that AHTI, besides reducing fasting glucose, can reduce weight gain via food intake reduction
A trypsin Inhibitor from tamarind reduces food intake and improves inflammatory status in rats with metabolic syndrome regardless of weight loss
(Nutrients, 2016-09) Maciel, Bruna Leal Lima; Carvalho, Fabiana Maria Coimbra de; Lima, Vanessa Cristina Oliveira de; Costa, Izael de Souza; Medeiros, Amanda Fernandes de; Serquiz, Alexandre Coelho; Lima, Maíra Conceição Jerônimo de Souza; Serquiz, Raphael Paschoal; Uchôa, Adriana Ferreira; Santos, Elizeu Antunes dos; Morais, Ana Heloneida de Araújo
Trypsin inhibitors are studied in a variety of models for their anti-obesity and anti-inflammatory bioactive properties. Our group has previously demonstrated the satietogenic effect of tamarind seed trypsin inhibitors (TTI) in eutrophic mouse models and anti-inflammatory effects of other trypsin inhibitors. In this study, we evaluated TTI effect upon satiety, biochemical and inflammatory parameters in an experimental model of metabolic syndrome (MetS). Three groups of n = 5 male Wistar rats with obesity-based MetS received for 10 days one of the following: (1) Cafeteria diet; (2) Cafeteria diet + TTI (25 mg/kg); and (3) Standard diet. TTI reduced food intake in animals with MetS. Nevertheless, weight gain was not different between studied groups. Dyslipidemia parameters were not different with the use of TTI, only the group receiving standard diet showed lower very low density lipoprotein (VLDL) and triglycerides (TG) (Kruskal–Wallis, p < 0.05). Interleukin-6 (IL-6) production did not differ between groups. Interestingly, tumor necrosis factor-alpha (TNF-α) was lower in animals receiving TTI. Our results corroborate the satietogenic effect of TTI in a MetS model. Furthermore, we showed that TTI added to a cafeteria diet may decrease inflammation regardless of weight loss. This puts TTI as a candidate for studies to test its effectiveness as an adjuvant in MetS treatment

SIGAA

Navegando por Autor "Serquiz, Raphael Paschoal"