Time Course Study of Erk1/2 Activity and Cell Viability in Lipopolysaccharide Challenged Pc12 Cells Publisher



Javadpour P^{1, 3} ; Askari S^{1, 3} ; Azizi F² ; Ghasemi R^{1, 3}
Source: Physiology and Pharmacology (Iran) Published:2021

Abstract

Introduction: Extracellular signal-regulated kinase 1 and 2 (ERK1/2) are important mem-bers of mitogen-activated protein kinases (MAPK), which are actively involved in the shap-ing of cellular responses to different stimuli; however, these responses are somehow con-tradicting. It is believed that time is a crucial factor in the determination of these effects. Therefore, the present work was designed to obtain a more vivid view about the effect of time on ERK activity and its relation to cell viability. Methods: In the first step, we challenged cultured PC12 cells with different doses of lipo-polysaccharides (LPS) for different time intervals (3, 6, 24 and 48h) and the cell viability was checked by MTT test. Thereafter, we cultured the cells in 6-well plates and treated them with the effective dose (10µg/ml) for the abovementioned intervals and the level of ERK phosphorylation, as the active form, was assessed in the Western blotting analysis. Results: The results showed that treating cells with 10µg/ml LPS reduces cell viability after 48h. While being ineffective in shorter periods of time, ERK activity has a fluctuating trend, so that it reaches the highest level at 6h, thereafter it declines to the lowest level at 24h and partially increases again at 48h. Conclusion: These results imply that time is a determinant factor in the activity of ERK and single-point assessments may result in misinterpretation. © 2021, Iranian Society of Physiology and Pharmacology. All rights reserved.