The effects of different concentrations of acetylsalicylic acid on proliferation and viability of lymphocytes in cell culture

Abstract

Numerous studies conducted on acetylsalicylic acid (ASA, aspirin) confi rmed that ASA inhibits proliferation and induces apoptosis in various types of human cells. Therefore, it was of interest to examine possible eff ects of different concentrations of ASA on viability and proliferation of lymphocytes in the cell culture. After separation from blood, lymphocytes were suspended in RPMI 1640 medium and cultured at 37°C. Solution of ASA was added to cultures after 24h, in fi nal concentrations of 1, 3 and 5 mmol/l. After 48h, proliferative response was evaluated by WST-1 assay. Signifi cant diff erence in viability between controls and cell cultures treated with ASA in three different concentrations was observed (p < 0,01). Percents of viable cells in cultures after application of 1,3 and 5 mmol/l ASA were 9,9%, 2,5% and 16,9% (compared to controls), respectively. To determine whether this cytotoxic eff ect was result of induction of apoptosis, DNA from cell cultures was isolated and subjected to agarose gel electrophoresis. Fragmentation of DNA was not detected, excluding apoptosis as possible cause of cytotoxic eff ects. Addition of ASA caused change of initial extracellular pH value for each treated culture. After addition of 1 mmol/l ASA, pH of culture was 7,19, after 3 mmol/L, 6,99 and after addition of 5 mmol/l solution, pH was 6,75. Decreased lymphocyte viability could be attributed to either the eff ects of the added substance or possible further acidifi cation of cell cultures during three days of incubation.