SILVER NANOPARTICLES Ag(0) CAUSE APOPTOSIS OF MOUSE MACROPHAGES IN VITRO INFECTED WITH BCG VACCINE WITHOUT INDUCTION OF OXIDATIVE STRESS IN CYTOPLASM AND MITOCHONDRIA

Original title

НАНОЧАСТИЦЫ МЕТАЛЛИЧЕСКОГО СЕРЕБРА Ag(0) ВЫЗЫВАЮТ АПОПТОЗ МАКРОФАГОВ МЫШЕЙ, IN VITRO ИНФИЦИРОВАННЫХ МИКОБАКТЕРИЯМИ ВАКЦИНЫ БЦЖ, БЕЗ ИНДУКЦИИ ОКИСЛИТЕЛЬНОГО СТРЕССА В ЦИТОПЛАЗМЕ И МИТОХОНДРИЯХ

Authors

Elena Ufimtseva

Contact information

Research Institute of Biochemistry, Federal Research Center of Fundamental and Translation Medicine, Novosibirsk, Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.

Pages

142-144

DOI

10.31255/978-5-94797-318-1-142-144

Abstract

Tuberculosis (TB) caused by Mycobacterium tuberculosis is currently an extremely widespread disease in the world, with high morbidity and mortality rates despite the significant advance in medicine. With the emergence and increase of M. tuberculosis resistant to multiple antibiotics, the development of new and effective antimicrobial reagents free of resistance is required for the successful treatment of TB infection. Due to their bactericidal properties, silver nanoparticles (Ag(0)-NPs) are the most frequently applied nanomaterials.

We have determined anti-mycobacterial activity of Ag(0)-NPs with different size 2-5, 10-15 and 35-40 nm at the 5.0 µg/ml concentration in mouse bone-marrow cells and peritoneal macrophages in vitro infected with the Bacillus Calmette-Guérin (BCG, a strain of attenuated live M. bovis) vaccine, because good concordance was found between the results on the sensitivity of M. bovis BCG and M. tuberculosis to diverse compounds. Then, we have studied the molecular mechanisms of Ag(0)-NPs action, such as ROS (reactive oxygen species) generation, apoptotic or necrotic death, and inflammation (the production of TNFα, IFNγ, and IL-1α), in BCG-infected mouse peritoneal macrophages with or without the addition of the antioxidant GSH (reduced glutathione), the mitochondrial antioxidant LiCl, the proapoptotic agent ABT 737, and the ROS inducer Н2О2. In a result, we revealed the activation of caspase 3/7 (CellEvent Caspase-3/7 Green Detection Reagent staining) and apoptotic death (Annexin V-FITC staining) in animal cells after 2, 4, and 6 hours of the Ag(0)-NPs treatment, but not after the exposure only to BCG mycobacteria or isoniazid. We did not observe the hydrogen peroxide (H2O2, H2DCFDA staining), the mitochondrial superoxide radical (O2−, MitoSOX Red indicator staining), and other ROS types (CellROX Deep Red Reagent staining) generation in the all investigated cell cultures for 2, 4, and 6 hours of the Ag(0)-NPs treatment. Also, we did not determine P53 protein stabilization and γH2AX, a phosphorylated version of the histone variant H2AX widely used as a marker for DNA double strand breaks very rapidly following exposure of cells to DNA damage, in mouse macrophages after the Ag(0)-NPs treatment.

After 24 hours of experiments, we detected the elevated ROS type production, but not H2O2 and mitochondrial O2−, in some BCG-infected mouse macrophages treated with the 35-40 nm Ag(0)-NPs, that was reduced by GSH treatment. We supposed this phenomenon was associated with the determined intensification of BCG mycobacteria killing in host macrophage lysosomes (Mycobacteria lipoarabinomannan and LysoTracker Red DND-99 dye fluorescent staining) at this time point.

In the hole, we did not found the activation of oxidative and genotoxic stress in mouse macrophages after exposure of cells with Ag(0)-NPs. Consequently, further studies of the exact mechanisms of Ag(0)-NPs bactericidal action in mouse macrophages with BCG mycobacteria are required.