Acrylamide treatment affects oxidative stress parameters in rat hepatocytes

Abstract

INTRODUCTION: Acrylamide (AA) is industrial toxic substance with neurotoxic and reprotoxic effects. AA is a Maillard reaction product formed during processing of starchy food at high temperature. OBJECTIVES: The objective of our study was to determine whether acrylamide treatment disturbs redox balance by altering nitrite, gluthatione (GSH), and malondialdehyde levels in rat hepatoma cell line (H4IIE). METHOD / DESIGN: Rat hepatoma cell line H4IIE was treated with 4 mM (IC20) and 4.5 mM (IC50) of AA for 24 h. The nitrite level in the medium was analyzed as an indicator of NO production following the Griess reaction method. After ultrasonic cell lysis in 2.5% sulfocalicylic acid, supernatant was analysed for the content of gluthatione. Lipid peroxidation was evaluated using thiobarbituric acid reactive substance assay (TBARS). RESULTS: Detected nitrite, malondialdehyde and GSH levels in rat hepatoma cell line H4IIE after acrylamide treatment are shown in Figure 1. Figure 1. Nitrite concentration (a), malondialdehyde (MDA) concentration (b), reduced glutathione (GSH) concentration (c) in H4IIE cells after treatment with 4 and 4.5 mM acrylamide (AA) for 24 h. Values in charts are means ± SEM of three experiments performed in triplicate. Mean values were significantly different from that of untreated control cells (*p<0.05). CONCLUSIONS: In rat hepatoma cell line H4IIE, exposure to AA caused significant concentration-dependent increase of nitrite level and lipid peroxidation (Fig. 1a, b). On the other hand, GSH content significantly decreased in a concentration-dependent manner in H4IIE cells (Fig.1c). Obtained results indicate that AA disturbs redox status in hepatocytes.