Avaliações da atividade antileucêmica em diferentes estratégias de produção de extratos da própolis âmbar

Abstract

Cancer is among the Brazil´s top three causes of death and is considered a public health problem, especially among developing countries, increasing interest in new active compounds. Apiculture products have gained attention due to their biological activities, mainly the propolis, which has been extensively studied demonstrating antifungal activity, antiviral, antibacterial, antioxidant and recently antitumor activity. The use of Brazilian propolis in "phytotherapy or apitherapy" should not be generalized due to the fact that its chemical composition is complex and highly variable according to geographic location, seasonal periods and plant distribution, being therefore of great importance the identification and isolation of active compounds from propolis extract to develop new drugs for cancer therapies. The objective of this work was to test different methods for the production of extracts of propolis from São Gabriel, denominated propolis Amber, evaluating its antileukemic activity through cell viability and IC50 test and to analyze the chromatographic profile of extracts by HPLC. The preparation of the hexane extracts and dichloromethane were carried out with the samples of the propolis of the city of São Gabriel / RS of the years 2014 and 2015. First, it carried out the maceration methodology of hot extraction by the Soxhlet extractor. After the obtained extracts were conducted the saponification reaction in basic methanolic medium being the product extracted obtaining only the unsaponifiable portion.Subsequently, in order to compare for biological activity, the maceration methodology was used, extracting the ambient temperature in the period of 7 days, with periodic agitations. The antileukemic activity was analyzed in the Jurkat cell line by the IC50 parameter and cell viability using Propidium Iodide exclusion test by flow cytometry and the chromatographic profile was analyzed by HPLC. The results showed antileukemic effect of the extracts of propolis in all extractions by the extraction methods and solvents with the exception of the unsaponified Hexane for both propolis samples. The IC 50 dosis of the extracts that presented antileukemic activity were concentrations between 0.2 to 0.5 μg / mL. When the chromatographic profile was carried out, they showed a distinctive peak at approximately 30 minutes in which we hypothesized that it is a possible phytochemical marker for amber propolis; however, we did not obtain any evidence yet about what peak could explain the activity differences between the extracts produced. In conclusion, the Soxhlet extraction was the more efficient method because of the reduction of the extraction time, as well as the Dichloromethane solvent by the extraction of apolar compounds and of low polarity that showed greater similarity between the samples activities, presenting similar data to those obtained with the ethanolic extract of this propolis.