Phytochemistry characterization of hexane extracts from branches and leaves of Endlecheria paniculata and inflammatory studies by sponge implants

The search for new anti-inflammatory drugs is one of the most explored areas of research in natural products, as several isolated molecules of plants have been shown to have promising effects in the treatment of this pathology. Endlecheria paniculata is a species of Lauraceae family, than in Brazil is native to the Amazon Rainforest, the Atlantic Forests and the Cerrrado1. Previous studies with E. paniculata identified some neolignans in the extracts from branches and leaves2. This class of compounds has displayed potent NF-κB (Nuclear Factor Kappa B) inhibition3, therefore presenting possible anti-inflammatory effects. This work presents the phytochemistry and anti-inflammatory activity of hexane extracts from branches and leaves of E. paniculata. Leaves and branches were collected in the ecological park of Pereque, in Cubatão-SP. Hexane extract from dried leaves (10.2 g) and branches (0.909 g) were prepared by maceration at room temperature from 224 g and 116 g, respectively. Extracts were analyzed by HPLC (High Performance Liquid Chromatography) and it was observed two compounds as majority. The isolation of major compounds by HPLC-preparative and characterization by NMR (Nuclear Magnetic Resonance) and HRMS (High Resolution to Mass Spectrometrywith electrospray ionization and positive mode analysis) it was identified two neolignans2. All hexane extracts were also analyzed by GC-MS (Gas Chromatography coupled Mass Spectrometry) and were isolated and identified: β-sitosterol4, stigmasterol, campesterol and palmitic acid. The anti-inflammatory effects of hexane extract from branches and leaves of E. paniculata was evaluated by a chronic inflammatory response induced by a murine implant sponge model for nine days5,6* (Ethics Committee of Animal Experimentation of the Federal University of Uberlândia, Minas Gerais CEUA 083/17). The inflammatory process was evaluable by measuring tissue levels of NAG (N-acetyl-β-D-glucosaminidase) and MPO (myeloperoxidase) activity, associated with macrophages and neutrophils accumulation in sponge induced fibrovascular tissue. The mouse treatment with hexane extracts of branches at 100 and 1000 ng doses increased MPO and NAG levels when compared with the control group (mouse treatment with PBS). The treatment with hexane extract of leaves at 10 ng dose reduced MPO level and at 100 and 1000 ng doses increased NAG. Besides that, in histological sections stained with toluidine blue a greater number of mast cells was observed in hexane extracts of branches at 10, 100 and 1000 ng doses and a reduction of this cell type was observed in hexane extracts of leaves in all doses tested.Thus, the data together suggest the hexane extract of leaves showed anti-inflammatory activity while the hexane extract of branches showed pro-inflammatory activity. We also analyzed the extracellular matrix deposition. Both extracts did not modify the soluble collagen production, but when total collagen deposited in histological sections stained with picrosirius red was analyzed, hexane extracts of branches at all doses showed decrease level and hexane extracts of leaves at 10, 100 and 1000 ng doses showed increase level. Thus, the treatment with hexane extract of branches showed antifibrogenic effect, while hexane extract of leaves showed fibrogenic effect.