ABSTRACT
Extracts of Serjania lethalis A. St.-Hil leaves and stems were tested in order to identify potential agents against Leishmania amazonensis. The hexane fraction (HF) and dichloromethane subfractions (DDF and MDF) showed leishmanicidal effect. The anti-promastigote IC50 values were 10.29 (HF), 11.41 (DDF) and 28.33µg/mL (MDF); whereas those against amastigote were 7.2 (HF), 8.1 (DDF) and 6.5µg/mL (MDF). Among the fractions and subfractions assayed, only HF altered the cell cycle of the parasite, increasing 3-fold the number of cells in the sub-G0/G1 phase. HF also changed the parasite mitochondrial membrane potential (ΔΨm) and the percentage of annexin-V-propidium iodide positive promastigotes. Our evaluations of the IC50 values showed that HF, DDF and MDF decreased NO production in infected macrophages stimulated with IFN-γ and LPS. Moreover, HF increased the production of TNF-α in Leishmania infected macrophages. This paper reports for the first time the leishmanicidal activity of extracts and fractions of Serjania lethalis leaves and also characterizes its leishmanicidal and immunomodulatory properties.
Subject(s)
Antiprotozoal Agents/pharmacology , Leishmania mexicana/drug effects , Macrophages, Peritoneal/drug effects , Magnoliopsida/chemistry , Plant Extracts/pharmacology , Animals , Annexin A5/analysis , G1 Phase/drug effects , Hexanes/chemistry , Immunomodulation , Inhibitory Concentration 50 , Interferon-gamma/immunology , Leishmania mexicana/growth & development , Leishmania mexicana/physiology , Lipopolysaccharides/immunology , Macrophages, Peritoneal/immunology , Macrophages, Peritoneal/parasitology , Membrane Potential, Mitochondrial/drug effects , Methylene Chloride/chemistry , Mice , Nitric Oxide/biosynthesis , Plant Extracts/chemistry , Plant Leaves/chemistry , Resting Phase, Cell Cycle/drug effects , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
An automated headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed in order to quantify a large number of volatile compounds in wines such as alcohols, ester, norisoprenoids and terpenes. The procedures were optimized for SPME fiber selection, pre-incubation temperature and time, extraction temperature and time, and salt addition. A central composite experimental design was used in the optimization of the extraction conditions. The volatile compounds showed optimal extraction using a DVB/CAR/PDMS fiber, incubation of 5 ml of wine with 2g NaCl at 45 °C during 5 min, and subsequent extraction of 30 min at the same temperature. The method allowed the identification of 64 volatile compounds. Afterwards, the method was validated successfully for the most significant compounds and was applied to study the volatile composition of different white wines.
