Cytotoxic activity induced by the alkaloid extract from Ipomoea carnea on primary murine mixed glial cultures

CABRERA A; PISTÁN M; TORRES ANA M; BUSTILLO S; CHOLICH, L

Jornada; XXIII Jornada Anual SAB 2021; 2021

Genus Prosopis spp., commonly known as ?algarrobo?, is a tree found in different regions of the world. In Argentina, P. flexuosa are geographically distributed throughout different ecoregions, including Chaco, Espinal Norte and Pampeana. Prosopis pods are among the ancient food resources used for animal feed, due to their nutritional value. Although its consumption is quite widespread, in Brazil neurological symptoms have been observed in animals associated with long-term intake of pods of another species of this plant, P. juliflora. In Argentina, intoxications with P. flexuosa have been reported in goats and cattle. Astrogliosis is considered an early marker of neurotoxicity as it is generally detected before any other toxic effects on neurons. This could be associated with the presence of the juliprosine and juliprosopine alkaloids previously identified in the total extract of P. flexuosa pods by HPLC-HRMS. The aim of the present work was to determine the cytotoxicity on glial cells of alkaloid enriched extracts of P. flexuosa collected in three different years, to evaluate the stability/variability of secondary metabolites present. Briefly, C6 Glioma cells (ATCC:CCL-107?) were resuspended and seeded in 96-well microplates, at initial density of 3.0-3.5 x 104cells per well in growth medium (DMEM-SFB 10%). When monolayers reached 80% confluence, different concentrations (10-50 µg/ml) of P. flexuosa extracts collected in the years 2019, 2020 and 2021 were added to the cells (200 μL/well). After 48 hours of incubation at 37°C and 5% CO2, cell viability was quantified by crystal violet staining. Results showed that P. flexuosa extracts from the three years studied, decreased cell viability in a dose-dependent manner. However, the extracts obtained from the years 2019 and 2020 proved to be more toxic (50 g /mL: 74% and 76% cytotoxicity respectively) than those from 2021 (50 g /mL: 36% cytotoxicity). These results show a possible variation in the stability and /or concentration of the alkaloid components present in Prosopis flexuosa pods according to the year of collection.