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Avaliação do efeito da fração lipídica extraída de Agaricus brasiliensis antioxidante e imunomoduladora in vitro e em modelo de sepse letal em murino
(Universidade Federal do Pará, 2021-01) LIMA, Kely Campos Navegantes; MONTEIRO, Marta Chagas; http://lattes.cnpq.br/6710783324317390; https://orcid.org/0000-0002-3328-5650
Sepsis is defined as a potentially fatal organ dysfunction caused by a dysregulated host immune response to an infection. During sepsis, dysregulation of the host response occurs with the excessive release of pro-inflammatory mediators, generation of reactive species with depletion of antioxidant defenses and cellular damage. As a result, the patient develops organ dysfunction. In this context, our group proposes the A.brasiliensis Lipid Fraction (FLAb) as a possible therapy for sepsis considering its immunomodulatory and systemic antioxidant activity in a murine sepsis model. Thus, the present study aimed to evaluate the activity of FLAb isolated in vitro and to evaluate the effect of treatment with FLAb alone or associated with the antibiotic ertapenem (F-Erta) on coagulation, antioxidant and immunomodulatory parameters in the lethal sepsis model in murine. For this, FLAb was kindly provided by Dr. Herta Dalla- Santa from UNICENTRO. In the present study, the antioxidant capacity of different concentrations of FLAb (1.25 and 5 μg/mL) was evaluated and in a RAW 264.7-Luc macrophage cell line, cytotoxicity, phagocytic capacity, nitric oxide, NF-κB activity and cytokines TNF-α and IL-6 were evaluated.The survival rates were analyzed 7 days in a model of CLP sepsis in swiss albino mice (Mus musculus), and treated with CLP+Salt (0.9%), CLP+FLAb (0.2mg/Kg), CLP+F-erta (0.2mg) /Kg; 30mg/Kg). For evaluation of on coagulation, antioxidant and immunomodulatory parameters, the mice were treated by 6 and/or 24h after CLP. In vitro, FLAb show antioxidant and anti-inflammatory activity in both concentrations. In vivo, all CLP+Salt animals died within a maximum of 48 hours while the FLAb and F-Erta treated groups survived the 7 days. During this period, clinical parameters of these animals were evaluated, the septic animals treated with saline showed piloerection, with little active level of consciousness and most of the time they were stopped in the cage, some of them had ocular secretion. In addition, animals treated with saline showed significant weight loss, reduced water and feed consumption resulting in death. The FLAb and F-Erta groups were active, with normal appearance, with normal breathing and heart rate, in addition to consuming water and food within normal limits. In the inflammatory site, peritoneal cavity, the treatment with FLAb showed an anti-inflammatory effect, decreased reactive oxygen species (ROS) and increased GSH antioxidant activity and protected from cell damage, maintaining neutrophil recruitment and nitric oxide levels (NO), reducing the bacterial load. Regarding coagulation parameters (platelet count, tp and ttpa), treatment with FLAb and F-Erta eliminated the bacterial load and protected the animals from tissue damage. In the liver, 6 hours after CLP the treatment with FLAb and F-ERTA was observed in the biochemical parameters protective effect, in addition, it presented immunomodulatory, antimicrobial and antioxidant activity preventing liver damage. In the parameters evaluated in the heart, the treatment with FLAb and F-ERTA after CLP protected the animals from cardiac damage through immunomodulatory, antimicrobial and antioxidant activity. In this sense, FLAb alone showed promise as a treatment and/or adjunct in sepsis, in addition to preventing organic dysfunction in septic animals.