Efeito antialérgico e imunomodulador dos alcaloides warifteina e metilwarifteina em modelo murino de Síndrome da Asma e Rinite Alérgicas Combinadas (CARAS)

Abstract

Combined Allergic Asthma and Rhinitis Syndrome (CARAS) is characterized by chronic inflammation of upper and lower airway and type 2 adaptive cellular immune response (TH2), with serum IgE-allergen-specific elevation, eosinophilic infiltration, and hyperplasia and hypertrophy of goblet cells. The pharmacotherapy of CARAS is based on active drugs that have a wide range of relevant side and adverse effects. Thus, the aiming at the search for new therapeutic alternatives, we evaluated the possible effects of treatment with warifteine or methylwarifteine, Cissampelos sympodialis alkaloids, a plant with anti-inflammatory potential, in murine model of ovalbumin (OVA)-induced CARAS. The experimental procedures were approved by the Animal Use Ethics Committee of UFPB (CEUA No. 2518050618) and, the data were analyzed by ANOVA one-way with statistical significance of p<0.05. BALB/c mice were separated into groups (n = 5) with normal animals (Basal); sensitized and challenged with OVA (OVA) animals; sensitized, challenged and treated intranasally with warifteine (War 2 mg/kg), methylwarifteine (Mwar 2 mg / kg) or budesonide (BUD 0.9 mg / kg) animals after the last allergen challenge. Animals treated with warifteine or methylwarifteine showed a decrease (p<0.05) in clinical signs, sneezing and nasal friction, and reduction (p <0.05) in histamine-induced nasal hyperreactivity from the 1nmol dose. Nasal lavage fluid (NALF) and Bronchoalveolar lavage fluid (BALF) cells from the different experimental groups were quantified and there was a reduction (p<0.05) in the number of total and differential cells, main eosinophils in the groups in warifteine or methylwarifteine treatments when compared to the OVA group. In the systemic context, only methylwarifteine reduced eosinophilia. Serum levels of OVA-specific IgE were reduced (p<0.05) in the groups treated with both alkaloids. Histological analysis revealed that the War and Mwar groups showed decreased infiltration of inflammatory cells in the subepithelial and perivascular region in the nasal cavity and the peribronchiolar and perivascular region in hematoxylin-eosin-stained lung tissue. Nasal and pulmonary goblet cell hyperplasia and hypertrophy, highlighted after PAS staining, were reduced in the War and Mwar groups. Alkaloids did not change the number of mast cells in the nasal and pulmonary tissues after Toluidine Blue staining. In contrast, the number of collagen fibers in the War and Mwar groups observed after staining with Gomori's Trichome was reduced. Besides, treatments with warifteine or methylwarifteine decreased (p<0.05) the cytokine levels of TH2 (IL-4, IL-13 and IL-5) and TH17 (IL-17A) profiles with increased levels of IFN-γ (TH1 profile), indicating an immunomodulatory effect of both alkaloids towards the TH1 profile over the TH2 profile. The expression of the transcription factor NF-кB was analyzed in granulocytes and lymphocytes of BALF and we observed that the groups War and Mwar showed reduction in NF-кB expression in both populations. The integration of pathways that favor the transcription of TH2 profile cytokines via NF-кB may underlie the alteration of the expression of this factor in granulocytes and lymphocytes.These data suggest that warifteine and methylwarifteine have antiallergic and immunomodulatory effects in CARAS murine model.