Allicin exert a protective effect on injured BV2 cells through AT1-Hsp70-iNOS cross-talk modulation.

MARÍA BELÉN RUIZ-ROSO; ALEJANDRA CAMARGO; FERES MOCAYAR; CAROLINA TORRES; LUCIANA MAZZEI; WALTER MANUCHA

Mendoza

Jornada; XXXVI Reunión científica anual de la sociedad de biología de cuyo; 2018

Sociedad de Biología de Cuyo

Lately, there are a lot of reports on the use of phytochemicals and their implications for human health as the neuroprotection effect of allicin (Alli) -an organosulfur compound from garlic- linked to a reduction of the oxidative stress, inflammation, and apoptosis. Oxidative stress and mitochondrial dysfunctions are common features leading to neuronal death in cerebral ischemia, traumatic brain injury, Parkinson's disease, Huntington's disease, and Alzheimer's disease; where the nitric oxide (NO) is a predominant effector of neurodegeneration. Parallel, heat shock protein 70 (Hsp70) upregulation may protect depression by downregulation of inducible nitric oxide synthase (iNOS) expression. To highlight, Alli attenuated loss of cell viability and neuronal apoptosis by iNOS decrease expression with an increased in the Hsp70 protein. Besides, Alli exerts protection like to losartan, and in silico analyses suggested that Alli and losartan could have a common mechanism involving AT1 interaction. AT1 overexpression is an early deleterious factor in many brain diseases. Aim: To evaluated if Alli may exert a critical anti-apoptotic/anti-inflammatory/anti-oxidative role in a model of neuronal injury, a murine microglial cell line (BV-2 cells), through AT1-Hsp70-iNOS axis modulation. To highlight, we also assessed if there is a possible protein's interaction related to BV-2 injury as well as the Alli improvement. Methods: BV-2, were cultured and after 80% confluence, cells were placed in quiescence for 24 hours. Immediately, BV-2 cells were injured with lipopolysaccharides (LPS, 100 ng/mL) and treated or not with Alli (50 μM) for 72h. The culture medium containing the LPS, Alli or both was replaced every 24 hours. All treatments were done in triplicate, and all experiments were replicated five to ten times. Cells were collected to perform the following determinations: Cell viability (MTT), pro-inflammatory cytokine levels, cellular oxidative stress (DHE), mitochondria isolation, NADPH oxidase activity (cells and mitochondrial fractions), iNOS, Hsp70 and AT1 expression (WB), nitrite levels (Griess), and protein-protein interactions by immunoprecipitation technique. Results: LPS in BV-2 cells provoked morphological changes, reduction in mean cell viability (p