Banca de DEFESA: Vanessa da Silva Eschimith

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : Vanessa da Silva Eschimith
DATE: 30/10/2023
TIME: 14:00
LOCAL: Plataforma Teams
TITLE:

EVALUATION OF AN ALKALOID FROM Pilocarpus microphyllus ON THE FUNCTION OF IMMORTALISED MICROGLIA

KEY WORDS:

Mitochondrial activity. EROS and ERNS. Phagocytic capacity. Lipid corpuscle. Cytokines.

PAGES: 85
BIG AREA: Outra
AREA: Tecnologia e Inovação
SUMMARY:

Introduction: The search for new molecules is important to counteract the large number of infectious agents, diseases and conditions through the mechanisms of drug resistance and tolerance. Alkaloids are a broad group of molecules whose biological activity has been increasingly explored due to their ability to interact with cell receptors and proteins. Epiisopilosin (EPIIS) obtained from the species Pilocarpus microphyllus has low cytotoxicity and anti-inflammatory effects. Considering the function of macrophages in the immune system, this study evaluated the effect of different concentrations of EPIIS on the innate function of immortalized macrophages/microglia. Material and Methods: This is an experimental, descriptive and in vitro study carried out with C20 (human microglia) and BV2 (murine microglia) cells to evaluate the effect of different concentrations of EPIIS on cell viability (MTT method and chemiluminescence), cell adherence on coverslips, and by flow cytometry, the production of reactive oxygen (EROS) and nitrogen (ERNS) species, the production of lipid corpuscles (LC) and cytokines; by morphological method, the phagocytic index (PI) and the production of the superoxide radical were assessed (O₂^º). Results: 1) the different concentrations (0.5 to 128 µg/mL) did not affect the mitochondrial metabolism of C-20 cells when incubated for 24 h (MTT test); 2) chemiluminescence showed that 1 or 4 at 32 µg/mL increased the mitochondrial activity/viability of C-20 cells, but reduced it with 128 µg/mL; 3) incubation of BV-2 cells with 0.5 µg/mL/2h or with 32, 64 or 128 µg/mL for 24 h increased the % of viable cells (MTT test), but the other concentrations did not affect the mitochondrial metabolism of the cells (2 or 24 h); 4) there was a reduction in the % of adhered C-20 cells by incubation for 24 h with 4.0 or 64.0 µg/mL; 5) the highest concentration (64.0 µg/mL) increased the stimulated or basal production of EROS and the basal production of ERNS by C-20 cells; 6) the stimulated production of ERNS (LPS or LPS + TNF) was inhibited by C-20 cells with 64.0 µg/mL; 7) the basal production of CL was higher in C-20 cells with 0.5 µg/mL, but the stimulated one (LPS) was inhibited with 64.0 µg/mL; 8) the lowest concentration (0.5 µg/mL) increased the basal production of IL-17A in C-20 cells, but was inhibited in the presence of LPS at 0.5, 4.0 or 64.0 µg/mL; 9) the three concentrations of EPIIS did not affect the basal production of the cytokines IL-2, IL-6, NTF, INF, IL-4 or IL-10 in C-20 cells; 10) 4 µg/mL/2h increased the IF of BV-2 cells by the highest % of phagocytizing cells (%cels), while 0.5 or 4.0 µg/mL for 24 h reduced the IF by the lowest %cels and the lowest number of yeasts ingested (Mlev). As for the incubation time (2 or 24 hours) with BV-2 cells, the results showed that: 11) there was a reduction in the production of the O2º radical at the three concentrations at 2 h, but not at 24 h; 12) compared to 2 h, there was an increase in Mlev at 64 µg/mL/24 h; 13) the IF was higher at 24 h in the control, than at 2 h, due to the higher %cels and Mlev; 14) the production of the O2º radical was higher at 64 µg/mL/24 h, compared to 2 h. Conclusion: All the functional tests showed that the three concentrations of EPIIS affected the mitochondrial activity of microglia, the expression of adhesion receptors, the production of EROS and ERNS, the production of lipid corpuscles/cytokines and phagocytic capacity, demonstrating the potential of EPIIS to modulate the innate immune response of microglia, thus suggesting the continuation of studies aimed at future biomedical applications in vivo.

COMMITTEE MEMBERS:
Presidente - 3424061 - SELMA APARECIDA SOUZA KUCKELHAUS
Interno - 2243078 - LUIZ ANTONIO SOARES ROMEIRO
Externa à Instituição - HELIARA MARIA SPINA CANELA
Externa à Instituição - THAIS TAMARA CASTRO E SOUZA MINUZZI