RESUMO
Plasma membrane anchored nucleotidases (E-ATPDases), as the E-NTPDase family, could hydrolyze and regulate the pericellular levels of nucleotides in lymphocytes. Each immune organ has a different microenvironment and display lymphocytes with different functions and phenotypes. Considering the different functions of each resident subpopulations of lymphocytes, the E-ATPDases activities in bone marrow (BML), thymus (TL) and mesenteric lymph node (MLL) lymphocytes of Wistar rats were characterized. The hydrolysis of extracellular nucleotides (eATP and eADP) showed linearity in time of reaction between 0 and 120 min, and concentration of lymphocytes expressed in proteins between 1 and 6 µg protein in the reaction medium. The optimal activity was attained at 37 °C in a pH value of 8.0. The necessity of the cofactors Ca2+ and Mg2+ for the enzymatic activity was confirmed through a curve of concentration of 0-1000 µM in the reaction medium, with both cofactors showing similar effects in the enzymatic activity. The Chevillard plot revealed that the hydrolysis of eATP and eADP occurred at the same active site of the enzyme. The analyses of E-ATPDases inhibitor and enzyme specificity showed possible involvement of E-NTPDase isoforms - 1 and - 2 in the isolated cells. Furthermore, different kinetic behavior of the nucleotide hydrolysis in each resident subpopulation lymphocyte was observed in this study, as MLL showed the higher Vmax with the lowest km values, while TL had the lowest Vmax and high km values. The kinetic values for E-NTPDase activity of each immune tissue lymphocytes can be an important therapeutic target for numeral immune-related diseases.
RESUMO
Metastatic melanoma is a very aggressive skin cancer. Platelets are constituents of the tumor microenvironment and, when activated, contribute to cancer progression, especially metastasis and inflammation. P2Y12 is an adenosine diphosphate receptor that triggers platelet activation. Inhibition of P2Y12 by clopidogrel bisulfate (CB) decreases platelet activation, which is also controlled by the extracellular concentration and the metabolism of purines by purinergic enzymes. We evaluated the effects of CB on the viability and proliferation of cultured B16-F10 cells. We also used a metastatic melanoma model with C57BL-6 mice to evaluate cancer development and purine metabolism modulation in platelets. B16-F10 cells were administered intraperitoneally to the mice. Two days later, the animals underwent a 12-day treatment with CB (30 mg/kg by gavage). We have found that CB reduced cell viability and proliferation in B16-F10 culture in 72 h at concentrations above 30 µm. In vivo, CB decreased tumor nodule counts and lactate dehydrogenase levels and increased platelet purine metabolism. Our results showed that CB has significant effects on melanoma progression.
Assuntos
Melanoma Experimental , Melanoma , Neoplasias Cutâneas , Animais , Camundongos , Clopidogrel/farmacologia , Modelos Animais de Doenças , Camundongos Endogâmicos C57BL , Melanoma Experimental/tratamento farmacológico , Microambiente TumoralRESUMO
OBJECTIVES: This study was aimed at assessing the anti-arthritic effects of hesperidin on the inflammatory markers in serum/plasma, ectoenzymes activity in platelet, reactive oxygen species (ROS), apoptosis and cell cycle in bone marrow cells of a rat model of arthritis. METHODS: Fifty-six adult female Wistar rats (245-274 g) were grouped into eight of seven rats each: control rats given normal saline or 40 mg/kg of hesperidin or 80 mg/kg of hesperidin, 0.2 mg/kg of dexamethasone, arthritic rats given normal saline, or 40 mg/kg of hesperidin or 80 mg/kg of hesperidin, and 0.2 mg/kg of dexamethasone. Myeloperoxidase and nitrate plus nitrite levels were evaluated in the plasma and serum, respectively. The ecto-nucleoside triphosphate diphosphohydrolases, ecto-5'-nucleotidase and ecto-adenosine deaminase activities were assessed in platelets. Subsequently, the cells of the bone marrow were obtained, and the assays for ROS, apoptosis and cell cycle were evaluated using flow cytometry. KEY FINDINGS: The results showed that hesperidin mitigated inflammation, modulated adenosine nucleotides and nucleoside hydrolysing enzymes and levels, minimized ROS intracellularly, attenuated apoptotic process and activated cell cycle arrest in arthritic rat. CONCLUSION: This study suggests that hesperidin could be a natural and promising anti-inflammatory compound for the management of arthritis.
Assuntos
Anti-Inflamatórios/farmacologia , Artrite Experimental , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Citrus/química , Hesperidina/farmacologia , Hidrolases/metabolismo , Inflamação , 5'-Nucleotidase/metabolismo , Adenosina Desaminase/metabolismo , Animais , Anti-Inflamatórios/uso terapêutico , Apoptose , Artrite Experimental/tratamento farmacológico , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Feminino , Adjuvante de Freund , Hesperidina/uso terapêutico , Inflamação/metabolismo , Inflamação/prevenção & controle , Proteínas de Membrana/metabolismo , Nucleosídeo-Trifosfatase/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Pirofosfatases , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismoRESUMO
Hyperlipidemia generates deposition of lipids, inflammation, and oxidative damage in cells and tissues, including those of the brain. Tucumã (Astrocaryum aculeatum) fruits contain bioactive compounds with antioxidant and anti-inflammatory effects. We evaluated the action of Tucumã extract on memory and brain cortex redox balance in hyperlipidemic rats. For 30 days, Wistar rats received Tucumã extract (250 mg/kg). Then, hyperlipidemia was induced by intraperitoneal administration of Poloxamer-407. Twenty-four hours later, the object recognition index was measured. The animals were euthanized for sample collection 36 hr postinduction. Hyperlipidemic animals showed memory loss and an imbalance between reactive species and intrinsic antioxidants. We found that Tucumã prevented memory loss and protein and lipid oxidative damage and prompted a better antioxidant response in the cerebral cortex of rats with hyperlipidemia. These findings suggest a neuroprotective effect and nutraceutical potential of Tucumã. PRACTICAL APPLICATIONS: In the present work, we demonstrated that induced hyperlipidemia in rats caused memory loss and redox unbalance, both factors prevented by the administration of Tucumã (Astrocaryum aculeatum) extract. Two aims were fulfilled with these results. The first was to show that hyperlipidemia affected brain function through oxidative damage and concerned basic research. The second was to offer a therapy that prevented this harm and could be applied in the clinic. Tucumã has ethnopharmacological importance through the consumption of fruits or the administration of extracts and oils by a population that was shown to enjoy improved health and longevity. Here, we show evidence that Tucumã contributes to the maintenance of brain health by preventing memory loss and oxidative damage, a nutraceutical supplement that may aid the prevention of vascular, inflammatory, and brain diseases.