engleski

The establishment of the method for measurement of the markers of oxidative stress in HeLa cell culture

OBJECTIVE: The level of oxidative stress is determined by the balance between reactive oxygen species (ROS) production and their elimination by antioxidative systems, such as antioxidative enzymes and non-enzymatic antioxidants. Hyperbaric oxygen therapy (HBOT) is therapeutic modality in which a patient breathes 100% oxygen, while the pressure of the treatment chamber is increased to more than one atmospheric pressure. Oxygen, as reactive molecule can induce many enzymatic pathways. In whole animal experiments, acute exposure to HBOT has been shown to increase oxidative stress, while intermitent exposure have no such effect. The aim of this study was to establish a method to measure oxidative stress in cervical cancer cell line (HeLa) and RPMI 1640 medium exposed to acute hyperbaric oxygenation. DESIGN AND METHODS: cervical cancer line HeLa cells were cultured into cell culture flask (75 cm²) in RPMI 1640 medium with L-glutamine + 10% FBS + 1% Pen / Strep in a controlled atmosphere (37 ° C, 5% CO2, above 80% humidity). When confluence was reached 80 %, cells were exposed to an acute oxygen pressure increase (1x2 h at 2 atm) in the hyperbaric chamber (n=8). The control cells were exposed to room air of the same temperature and duration of the experiment (n=4). The level of oxidative stress in Hela cells and medium was determined (by measuring thiobarbituric acid reactive substances (TBARS) and antioxidative capacity by ferric reducing capacity of plasma (FRAP). Statistical analyses were performed using a Student's T-Test. Statistical significance level was set to p<0.05. RESULTS: The obtained results show there are no statistically significant differences between the controll group and group exposed to HBOT (cells p=0.295 TBARS, p=0.189 FRAP and medium p=0.985 TBARS, p=0.507 FRAP). CONCLUSIONS: Acute hyperbaric oxygenation did not significantly affect the level of oxidative stress in HeLa cell cultures.

HeLa cell culture ; oxidative stress

nije evidentirano