Efeitos celulares e comportamentais de peptídeos bioativos de baixo peso molecular extraídos de Phaseolus vulgaris
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2020-11-18
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Universidade Federal de Goiás
Resumo
INTRODUCTION: Approximately 3.000 tons of beans are not used in human consumption due to their hardening, which makes them a product of low commercial value. Several studies with bioactive peptides derived from beans have therapeutic potential to treat various diseases. OBJECTIVES: to evaluate the effects of a low molecular weight bean extract (Phaseolus vulgaris) hardened on: i) cytotoxic and cytoprotective effects on endothelial cells; ii) production of reactive oxygen species (ROS) and nitric oxide (NO) in endothelial cells; iii) oxidonitrergic-dependent vasodilator effects; iv) anxiety and depression behavior in rats; v) the antioxidant effect on rat brain. METHODS: The extract was composed of a peptide fraction less than 3 KDa (PV3) of the hardened bean residue. The PV3 sequences were corrected by mass spectrometry coupled to liquid chromatography and were analyzed with computational tools. Human umbilical vein endothelial cells (HUVEC) were treated with PV3 in the following procedures: 10 μg/ml, 20 μg/ml, 30 μg/ml and 250 μg/ml. Cellular oxidative stress was caused by 3% H2O2. Cytotoxicity and cytoprotective effects were obtained by the MTT assay, while ROS and NO were quantified by fluorescent probes (DHE and DAF-FM) by Confocal Microscopy. The vasodilator effects of NO3 and endothelium-dependent PV3 were obtained in the supplied aortic rings. The behavioral effects of acute and chronic PV3 treatments were adopted by three protocols: i) elevated plus maze test (EPM) to assess the effect of the anxiolytic type. ii) open field (OF) to assess locomotion and exploration; iii) forced swimming (NS) to test the behavior of the depressive type. The involvement of catecholaminergic pathways in the effects evoked by PV3 was tested using the enzyme tyrosine hydroxylase inhibitor, AMPT (200mg / kg). RESULTS: 35 peptides with an average mass of 1.14 KDa were identified. There was no cell death from treatment with PV3 10μg/mL and 20 μg/mL. PV3 30μg/mL increased cell viability, whereas cytotoxicity was observed only with PV3 250 μg/mL. Only PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased the release of NO without causing cell death. It was also able to reduce the relative production of cell ROS induced by H2O2. The vasodilator effects of PV3 were based on the release of NO dependent on the endothelium. In the EPM test, the acute injection of PV3 (50μg / kg) increased the frequency and the time spent in the open arms, suggesting an anxiolytic effect. In the OF test, PV3 (50μg / kg)
increased the frequency of crossings and immobility time, showing that PV3 does not impair locomotion, which corroborates the anxiolytic effect found in the ECL. In the FS test, PV3 (50μg / kg) reduced the immobility time, suggesting an effect similar to the antidepressant. The anxiolytic-type effect found after acute injections was absent in chronic treatment with PV3 (50μg/kg). AMPT was able to reverse the effect of the anxiolytic type and the antidepressant type evoked by acute PV3 (50μg/kg). CONCLUSION: PV3 has low cytotoxicity, the ability to reduce ROS and increase NO in the endothelium, in addition to promoting anxiolytic and antidepressant effects with catecholaminergic involvement.
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Phaseolus vulgaris , Feijão , Bioativo , Peptídeo bioativo , Citotoxicidade , Óxido nítrico , Espécie reativa de oxigênio , Estresse oxidativo , Vasodilatação , Comportamento , Ansiolítico , Antidepressivo , Bean , Bioactive , Bioactive peptide , Cytotoxicity , Nitric oxide , Reactive oxygen species , Oxidative stress , Vasodilation , Behavior , Anxiolytic , Antidepressant
Citação
GRAZIANI, D. Efeitos celulares e comportamentais de peptídeos bioativos de baixo peso molecular extraídos de Phaseolus vulgaris. 2020. 114 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Goiás, Goiânia, 2020.