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Modulation of the pharmacological, biochemical and enzymatic activities of basic sPLA2 from Bothrops jararacussu and Crotalus durissus ssp using purified extracts obtained from Tithonia diversifolia.

Grant number: 10/09988-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): September 01, 2010
Effective date (End): October 31, 2012
Field of knowledge:Biological Sciences - Pharmacology - Ethnopharmacology
Principal Investigator:Marcos Hikari Toyama
Grantee:Veronica Cristina Gomes Soares
Home Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

The objective of this project is to evaluate the inhibitory effect of extracts from the aerial parts of Tithonia diversifolia, on the activity of the venom and fractions of sPLA2 basic obtained from Bothrops jararacussu and Crotalus durissus ssp. In addition to testing on enzymes isolated from the venom of Crotalus durissus ssp and Bothrops jararacussu, tests will be performed on the enzyme activity of phospholipase sPLA2 obtained from bovine pancreas and intracellular sPLA2. To determine the possible anti inflammatory activity of these extracts, the basic sPLA2 are previously incubated with the extracts, then tested for: paw edema, skin edema, myotoxicity, neurotoxicity and aggregation. The poison effect will be determined after pre incubation of venom with total extracts at different concentrations, and the execution of these tests referred to the basic sPLA2. The anti hemorrhagic effects of these compounds on clotting time and plasma fibrinogen degradation induced by the poison total will also be evaluated. The activity of the extracts will be evaluated by enzyme assays and objective of this project will determine the cytotoxic effect, the same, on cell line of fibroblasts.The effects of these extracts on the basic structure of sPLA2 be evaluated by spectroscopic techniques of circular dichroism, fluorescence and UV-Vis scanning. The techniques of chromatography and HPLC molecular exclusion will be used to check any changes in the state of oligomerization of sPLA2 basic. The technique of reverse phase HPLC, will assess possible changes in hydrophobicity of sPLA2 basic. The completion of the chemical structure activity relationship of the main constituents of the extracts will be held by other specific protocols for analysis of amino acids, these techniques employed to characterize the structural changes in the basic sPLA2 by extracts from Tithonia diversifolia.

Academic Publications
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