Avaliação da atividade anti-inflamatória e antifúngica in vitro e ex vivo da própolis verde brasileira frente a fungos do gênero Paracoccidioides spp.

Abstract

Paracoccidioidomycosis (PCM), a systemic, granulomatous mycosis, is caused by fungi of the genus Paracoccidioides spp., with two pathogenic species, Paracoccidioides brasiliensis (Pb18) and Paracoccidioides lutzii (Pl), which are thermally dimorphic, varying in mycelial and yeast. PCM has severe clinical consequences in its sufferers that require more extensive therapeutic options, given that there are a reduced number of medications and those available have a prolonged treatment time, therefore, complementary treatment is sought as a way of immunomodulating the immune system. It is known that patients with PCM have a deficient immune response to invasive pathogenic microorganisms, for this reason, a combination of Brazilian Green Propolis (PVB) is proposed, the aim of which is to modulate the immune system. Propolis has been mentioned in the literature due to its anti-inflammatory, antimicrobial and immunomodulatory activities. Objective: to study the anti-inflammatory and antifungal properties of PVB extract against the general course of PCM, seeking to find a reduction in infection levels and an antiPb18 and Pl response. Methodology: Initially, in vitro studies were carried out. The antimicrobial activity test was carried out on strains Pb18 and Pl to evaluate whether PVB showed microbicidal activity at a concentration of 500 mg/mL. Subsequently, splenic cells from the spleen of Swiss mice were used to evaluate the cytotoxicity of PVB at a concentration of 500 mg/mL. The animals were infected with fungal suspensions (Pb18 and Pl) using the “air pouch” technique, a type of subcutaneous air pocket, for 10 days. Treatment with PVB extract was carried out in the last three days of infection. At the end of the process, the cells were collected from the “air pouch” and separated into sediment and supernatant to carry out the following tests: quantification of nitric oxide, total proteins, catalase enzyme, peroxidase, cytokine dosage, cell viability, cell quantification and viable fungi and phagocytic activity of PMNs. Results: In in vitro results, PVB showed antimicrobial activity against both strains. In splenic cell culture, the PVB concentration tested did not show toxicity. When evaluating whether PVB improves the capacity of splenocytes to eliminate the fungus, regardless of the strain, the results of the PVB test were significant in relation to the control. Treatment in animals increased the viability of immune cells, total proteins, catalase, peroxidase and production of the cytokines INF-y and IL-12. Treatment with PVB was also able to reduce the number of viable fungi, when compared to the control groups (Pb18 and Pl), in addition to demonstrating greater phagocytic activity of PMNs. Conclusion: PVB was able to immunomodulate the immune system, since in the treated population there were higher levels of PMN activity, in addition, it reduced the number of viable fungi and stimulated the production of total proteins and cytokines linked to the type of response. Th1. In this sense, the application of PVB extract may be a promising factor throughout the treatment and outcome of mycosis.