Detalhes bibliográficos
Ano de defesa: |
2010 |
Autor(a) principal: |
Sousa, Fabiola Carine Monteiro de |
Orientador(a): |
Não Informado pela instituição |
Banca de defesa: |
Não Informado pela instituição |
Tipo de documento: |
Tese
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Tipo de acesso: |
Acesso aberto |
Idioma: |
por |
Instituição de defesa: |
Não Informado pela instituição
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Programa de Pós-Graduação: |
Não Informado pela instituição
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Departamento: |
Não Informado pela instituição
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País: |
Não Informado pela instituição
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Palavras-chave em Português: |
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Link de acesso: |
http://www.repositorio.ufc.br/handle/riufc/5536
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Resumo: |
Bothropoides erythromelas is responsible for a great deal of snakebites in Northeastern from Brazil. The venom of this snake induces acute renal failure. Isolated kidneys from Wistar rats, weighting 250 to 300g, were perfused with Krebs-Henseleit solution containing 6g% of bovine serum albumin for 120 min. The whole venom of Bothropoides erythromelas been previously studied (SOUSA, 2004) and used in this study for comparison with the treated groups with the PLA2 fractions Lys 49 and Asp 49 of the venom. The whole venom (10mg/mL) and the fractions PLA2 Lys 49 and Asp 49 of B. erythromelas (5mg/mL) were added into the system 30 min after the beginning of each experiment. The parameters studied included perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), percent sodium, potassium and chloride tubular transport (%TNa+, %TK+ and %TCl-), percent sodium, potassium and chloride proximal transport (%pTNa+, %pTK+ and %pTCl-), sodium, potassium and chloride excretion (ENa+, EK+ e ECl-) and osmotic clearance (Cosm) (p< 0.05*). The control group perfused with albumin was functionally stable for over 120 min. The infusion of venom caused a significant increase in UF, ENa+, ECl- and Cosm and a decreased in PP, RVR, %TNa+, %TK+, %TCl-, %pTNa+, %pTK+ and %pTCl-. The GFR and the EK+ decreased at 60 min and increased at 90 and 120 min when compared with control group. The infusion of Lys 49 caused a significant increase in PP, UF, ENa+, EK+ and Cosm and decreased the GFR and the %TNa+ when compared with control group. Lys49 did not modify the others functional kidney parameters. The infusion of Asp 49 only modify the functional kidney parameters %pTK+ (decreased) and EK+ (increase) when compared with control group. Lys 49 showed a similar effect at whole venom in parameters UF, %TNa+, ENa+, EK+ and Cosm and Asp 49 in parameters %pTK+ and EK+. The histological analysis showed a mild amount of a proteinaceous substance in the renal tubules and glomeruli of kidneys perfused with the venom, Lys 49 and Asp 49, as well as focal areas of apoptosis/necrosis in perfused kidneys with Lys 49 e Asp 49. MDCK cells were cultured in RPMI 1640 medium supplemented with 10% vv fetal bovine serum and then assessed in the presence of the whole venom, Lys 49 and Asp 49 of B. erythromelas in the concentrations (100; 50; 25; 12.5; 6.25 and 3.125mg/mL). The analysis of cytotoxic effects on MDCK cells was performed by MTT method. The venom promoted cytotoxic effect in the concentrations of 50 and 100mg/mL (IC50 =93.31mg/mL). Lys 49 promoted cytotoxic effect in the concentrations of 6.25; 12.5; 25; 50 and 100 mg/mL (IC50 = 38.29mg/mL). Asp 49 promoted cytotoxic effect in the concentrations of 50 and 100 mg/mL (IC50 = 158mg/mL). Also the levels of lactic dehydrogenase (LDH) were measured and no significant increase was observed with whole venom and Asp 49. Lys 49 promoted a significant increase in the levels of LDH only in the concentration of 100mg/mL. After culture of MDCK cells with the whole venom, at concentrations of 46.65 and 23.32µg/mL, was performed the real time polymerase chain reaction for evaluation of pro (Caspase-3, Caspase-8 and Bax) and antiapoptotic (Bcl-XL and Mcl-1) genes expression. The evaluation of pro and antiapoptotic genes expression with Lys 49 e Asp 49 did not realized. In the expression of pro-apoptotic genes the whole venom caused increase of caspase-3 at concentration of 23.32µg/mL and of caspase-8 at concentrations of 46.65 and 23.32µg/mL, when compared with negative (PBS) and positive (DOXO) controls and decreased the expression of Bax in both concentrations. In the expression of anti-apoptotic genes the whole venom caused significant induction of Mcl-1 only at a concentration of 46.65µg/mL and did not modify the expression of Bcl-XL, when compared with the negative control. The venom and the fractions PLA2 Lys 49 e Asp 49 of Bothropoides erythromelas is able to promote significant effects on renal function parameters and on MDCK cells, with indications of cell death by apoptosis through the extrinsic pathway. |