Study of the mechanisms of cell death due to exposure to ketamine, ethanol and the association of both substances in SH-SY5Y cells

• Grant number: 18/24796-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2019
• End date: June 30, 2020
• Field of knowledge: Health Sciences - Pharmacy - Toxicological Analysis
• Principal Investigator: Raphael Caio Tamborelli Garcia
• Grantee: Emidio Piñeiro Lopez Junior
• Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil

Abstract

The use of illicit drugs is a major public health problem, especially among adolescents and young adults. Ketamine, a drug with anesthetic properties, has been used illegally by health professionals because of its ease of production. Besides, the increasing consumption of alcohol in Brazil, by different age groups, especially in adolescence and early adulthood, is worrying. It is noteworthy that this is consumed along with other substances, such as new drugs, for example ketamine, and may cause irreversible damage to the central nervous system due to their association. Thus, the present study intends to investigate the mechanisms that permeate cell death after exposure to ketamine, ethanol and the association of both substances in SH-SY5Y human neuroblastoma cell culture. Two concentrations of ketamine and ethanol will be defined by means of concentration-response curves using cell viability assays (MTT): the highest concentration without toxic effect (NOAEL) and the lowest concentration in which there is a toxic effect (LOAEL). Subsequently, the pathways involved in cell death, from both isolated and associated groups, will be investigated using annexin/propidium iodide (flow cytometry). Also, proteins involved in the intrinsic (caspase-9) and extrinsic (caspase-8) pathway of apoptosis, as well as effector caspase (caspase-3) will be evaluated. Methods employed include: spectrophotometry, western blot and flow cytometry.