Commiphora leptophloeos (Mart.) J.B. Gillett (Burseraceae): estudo fitoquímico, toxicidade e avaliação do potencial antiinflamatório e antimicrobiano

Abstract

The specie Commiphora leptophloeos (Burseraceae) is a native plant from Brazil, belongs to caatinga biome and is known popularly as “imburana-de-espinho” and “imburana-decambão”. The ethnopharmacology studies mention the use of this specie in the treatment of inflammations and infections. In this context, the present study aimed to evaluate the toxicity, anti-inflammatory and antimicrobial activity in non-clinical in vitro and in vivo assays of leaf and stem bark extracts, and to isolate, identify and quantify chemical markers. In the phytochemical study, the hydroethanolic extract was prepared by maceration and the fractionation was performaded by liquid-liquid partition (dichloromethane, ethyl acetateAcOEt and n-buthanol-BuOH fractions). Isolation, structural elucidation and characterization of the extracts were performed by CPC, LC-MS, FIA-ESI-IT-MS/MS and 1H NMR and quantification of the content of markers in the extracts was made by HPLC-DAD and HPLCELSD. The toxicity of both extracts was evaluated by MTT and flow cytometry assays and in vivo by the acute toxicity test. The in vitro anti-inflammatory activity was evaluated using LPS-induced nitric oxide assay and in viv by carrageenan-induced paw edema and oral zymosan induced air pocket models. The antimicrobial activity was measured by the determination of minimum inhibitory concentration (MIC) and virtual screening of the isolated compounds was performed by in silico studies. From leaves extract, 2 flavonoids were isolated by CPC and 16 compounds were characterized by Mass spectrometry such as phenolic acids, glycosylated flavonoids derivatives of quercetin, luteolin and apigenin, and condensed tannins derivatives of catechin. From the stem bark extract, 2 tannins were isolated by CPC and 8 compounds were characterized as phenolic acids and procyanidins. The leaves extract at 200 µg/mL and stem bark extract at concentrations of 1, 10, 100 and 200 µg/mL demonstrated in vitro anti-inflammatory effect in the LPS-induced nitric oxide assay. In the carrageenan-induced paw edema of model, the extracts of leaves and stem bark at the doses of 100, 200 and 400 mg/kg, by oral route, significantly reduced the edema followed by reduction of myeloperoxidase (MPO) and in the zymosan-induced model of pouch-air, at doses tested, significantly reducing (p < 0.001), cell migration, total protein concentration, myeloperoxidase (MPO), and malondialdehyde (MDA) and the proinflammatory cytokine TNF-α and increased the production of the anti-inflammatory cytokine IL-10. In the evaluation of the antimicrobial activity, the extracts of the barks and the AcOET and BuOH fractions at concentrations of 20 to 100 μg/mL demonstrated a fungistatic and bactericidal effect. The in silico study indicated possible ligands of type B dimeric procyanidin and isovitexin related to anti-inflammatory and antimicrobial activity. The results obtained are unprecedent for the specie, justify its use in folk medicine and reveals that extracts of leaves and stem bark have a therapeutic potential for the development of herbal products with antiinflammatory and antimicrobial properties.