Estudo da espécie Echinodorus macrophyllus: investigação da eficácia e segurança com fins terapêuticos

Abstract

Millenary techniques are used to extract the pharmacological properties of plants. However, the indiscriminate use of these plants can pose risks to the population, making it necessary to have greater knowledge about this type of medicine. Brazil is a country rich in biodiversity and has various native plants with medicinal potential. The search for more knowledge about these plants is increasingly necessary to meet the demands of the population. This study aims to standardize the protocol for obtaining the tincture of Echinodorus macrophyllus (Kunth.) Mich., according to the Brazilian Pharmacopoeia, outlines its phytochemical profile using chromatographic techniques, and investigates its acute anti-inflammatory effect in vivo and its cytotoxic, genotoxic, and mutagenic potential in vitro. The tincture was obtained by macerating dried leaves of the plant in different concentrations of hydroalcoholic solution (55%, 60%, and 65%). The three samples yielded 7.1%, 7.4%, and 7.7%, respectively. UV-Vis spectrum scanning was performed to investigate the wavelength range where the substances present in the tincture predominantly absorb, with a maximum absorption observed at 342 nm. Chromatographic analyses were performed using thin-layer chromatography (TLC), revealing different classes of secondary metabolites. The NP-PEG revelation showed bands corresponding to specific markers of the Alismataceae family in the phenolic class. Still, the revelation with sulfuric vanillin did not show bands, suggesting the absence of terpenes. Analysis of the samples by high-performance liquid chromatography coupled with a diode-array detector (HPLC-DAD) showed similarity between the tested concentrations. The sample with the lowest alcohol content, T55, was selected for further assays. HPLC-DAD analysis with co-injection of chemosystematic standards from the Alismataceae family with the chosen sample revealed the presence of substances such as isoorientin, vitexin, isovitexin, and rutin. Given the need for further studies to ensure the safety and efficacy of therapeutic use of Echinodorus macrophyllus, MTT, WST-1, and LDH assays were conducted, which demonstrated the absence of cytotoxicity in Hep-G2 and HEK-293 cells. The in vitro micronucleus assay showed no genotoxicity, and the Ames test revealed no evidence of mutagenicity in the analyzed concentrations of the Echinodorus macrophyllus tincture. The paw edema and peritonitis experiments induced by carrageenan demonstrated the anti-inflammatory effect of doses of 0.25 and 2.5 mg/Kg of the tincture. Despite the results indicating the absence of cytotoxicity, genotoxicity, and mutagenicity, further analyses regarding the safety of the mother tincture of E. macrophyllus are necessary, in addition to a thorough investigation of its anti-inflammatory effect.