Effects of acrylamide and curcumin on cytotoxicity, genotoxicity and cell signaling pathways in leydig cells

The aim of this study is investigating the mechanism related to acrylamide induced toxic effects and possible therapeutic effects of curcumin on TM3 Leydig cells. Acrylamide treated cells were evaluated for cytotoxicity, reactive oxygen species (ROS), antioxidant related enzyme levels, apoptosis/necrosis rates, cytoskeletal damage, phosphorylation status of mitogen activated protein kinases (MAPKs) and genotoxicity. The possible ameliorative effect of curcumin administration was investigated. Acrylamide, which is widely used in industrial field, has been found to occur in foods prepared in high temperature in recent years. Due to the nutritional habits in the world and in our country, foods containing acrylamide are consumed a lot. Acrylamide is a compound formed by the reaction of carbohydrates and amino acids in foods with high temperatures (120°C). Particularly grain and starchy foods contain high amounts of acrylamide. In addition, acrylamide is widely used in the paper, textile and cosmetic industry and in water treatment system. In recent years, the use of medicinal plants to treat human diseases has become widespread. Curcumin, which is the active ingredient of the turmeric plant, is mostly used as a spice and food colorant, and characterized as a polyphenolic compound. Studies have shown that curcumin has antioxidant, antiinflammatory, antibacterial, anticarcinogenic and antimutogenic properties. TM3 Leydig cells were exposed to four different concentrations of acrylamide (1, 10, 100, 1000 µM) in the presence and absence of curcumin (2,5 µM) for 24 hours. Oxidative stress markers and antioxidant defense system parameters measured by spectrophotometric method, apoptosis and necrosis rates determining by propidium iodide-Hoechst stain and for mark the cytoskeletal proteins immunofluorescent staining was performed. Western blot analyzes were performed for determining the phosphorylation of MAPKs further Comet assay and micronucleus test applied for evaluate the genotoxicity. According to the results of our study, acrylamide causes detrimental effect on cell function of Leydig cell and damage to the cytoskeleton, it causes lipid peroxidation by triggering ROS formation and thus deteriorates antioxidant-oxidant balance in Leydig cells. Results also showed that acrylamide induces apoptosis and causes genotoxicity in Leydig cells associated with stimulating oxidative stress and activating MAPK pathway. Moreover, results showed that curcumin suppressed the detrimental effects of acrylamide. Taken together, we suggest that acrylamide, induced apoptosis through activation of MAPK signaling pathway and caused genotoxicity furthermore curcumin has a protective effect against the acrylamide induced damage in Leydig cells