Standardized phytoterapics for the treatment of chronic diseases: Cytotoxic, mutagenic and protective properties and the modulation of gene expression

Abstract

The program "BIOTA/FAPESP" aims to identify and characterize the biodiversity of the State of São Paulo, defining the mechanisms for its conservation, economic potential and sustainable use. Some preliminary studies considering the biological activities of plant extracts were performed in the first stage of the project, however, their registration requires the assessment of herbal extracts standardized according to international standards, which is the objective of this second step. Thus, this project aims to collaborate in this important thematic project through in vitro evaluation of cytotoxic and mutagenic properties of standardized extracts from different plant species of the cerrado of São Paulo. Cell lines employed will be gastric cells ACP02 (tumoral) and primary culture of gastric cells (normal line), since the extracts evaluated in this study are used in treatment of gastrointestinal disorders and diseases. To assess cell viability and the mutagenic potential of the extracts, it will be used, respectively, the MTT assay and cytokinesis-block micronucleus test. The combination of dyes acridine orange and ethidium bromide, as well as flow cytometry, will be used to evaluate apoptosis. In parallel, the protective activity of these extracts will also be evaluated against the DNA damage and cytotoxicity induced by benzo(a)pyrene, a potent mutagen. The extracts that present protective activity in cells of the normal strain, but not in the tumoral cells will also be investigated by quantitative polymeric chain reaction array (RT-qPCR array).Expression of genes involved in oxidative stress, antioxidant defense process and inflammation will be investigate to propose the mechanisms of action of the protective effects of these extracts.Keywords: apoptosis, cytotoxicity, gastric cells ACP02; mutagenicity, phytotherapics, RT-qPCR array,