Abstract
ABSTRACT Among the immune system cells, macrophages have an important role. Apamin, a bee venom constituent, is important in the defense of these insects. Thus, we aimed to evaluate the metabolism of J774 1.6 macrophage cell line when exposed to isolated and purified apamin, using cytotoxicity tests by MTT reduction and analysis by flow cytometry (apoptosis / necrosis, production of reactive oxygen species (ROS), membranous lipoperoxidation (LPO), electrical potential of the mitochondrial membrane (mMP) and DNA fragmentation). None of the tested concentrations (10 to 100μg/mL) were cytotoxic according to MTT reductions. Apoptosis rates decreased at concentrations of 2.5, 5.0, and 10.0μg/mL (P<0.05), while necrosis rates increased (P<0.05). However, rates of healthy cells at the highest tested concentration (10μg/mL) did not differ from control (P>0.05). Apamin did not alter ROS, LPO, or DNA fragmentation. Therefore, all analyzed concentrations (1.25 to 10μg/mL) decreased mMP. Such decrease in apoptosis might be due to a suppression of mitochondrial pro-apoptotic messengers, as this peptide causes no oxidative stress, lipid peroxidation, and DNA damage. Highly sensitive techniques are majorly important for proper interpretation of cellular toxicity mechanisms, combined with routine laboratory methods.
Highlights
Macrophages are important immune system cells that participate in innate immunity and help to regulate adaptive immune responses through phagocytosis and antigen-presentation to lymphocytes, and their production of mediators interfere with a wide range of cytokines (Geissmann et al, 2010)
None of the apamin concentrations tested here differed from the control cells, for the MTT reduction test, which remained in essential medium (E-MEM) medium throughout the entire experiment
A decrease in apoptosis rates with respective healthy cell rate reduction was observed when cells were exposed to the concentrations of 1.25, 2.5, and 5μg/mL apamin (P
Summary
Macrophages are important immune system cells that participate in innate immunity and help to regulate adaptive immune responses through phagocytosis and antigen-presentation to lymphocytes, and their production of mediators interfere with a wide range of cytokines (Geissmann et al, 2010). These cells originate from monocytes that migrate from blood vessels in response to antigenic stimulation (Van Ginderachter et al, 2006). Macrophages, when in tissues, mature and adapt to the environment, differentiating into several types of cells with specific function such as Kupffer cells in liver and microglia in the central nervous system (Gordon and Taylor, 2005). Apamin is a less known neurotoxin which acts as a calcium channel blocker in postsynaptic membranes of the central and peripheral nervous system, responsible for blocking the transmission of certain inhibitory impulses (Banks et al, 1979; Cruz-Landim and Abdalla, 2002)
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