Luteolin Protects Against Acrylamide-Induced Cellular Toxicity in Mouse Leydig Cells

Luteolin (Lut), a natural flavonoid, possesses the ability to protect the organism from reactive oxygen species (ROS) by modulating intracellular and extracellular signals. This study was conducted to investigate whether Lut exhibits a protective effect against acrylamide (Acr)-induced reproductive toxicity and to reveal the mechanisms underlying Leydig cell damage, a model cell in the male reproductive system. TM3 mouse Leydig cells were treated with Acr (1 mM) and/or Lut (1 and 5 μM) for 24 h. To determine the potential protective effects of Lut against the toxicity caused by Acr on Leydig cells, cell viability, combination index, and lactate dehydrogenase activity were examined for cytotoxicity. The effect of Lut on oxidative stress parameters in Acr-induced cellular damage was revealed by evaluating lipid peroxidation, total ROS, glutathione, and antioxidant enzymes (SOD, CAT, and GPx). In addition, we evaluated the expression levels of genes related to apoptosis (Trp53, Casp3, Bax, and Bcl2) and the apoptotic rate in the Acr and/or Lut groups. The findings indicated that Lut inhibited the cytotoxicity elevated by Acr in Leydig cells. The evaluation of the combination index in the groups treated with Lut and Acr together revealed that Lut exhibited an antagonistic effect. Furthermore, it was revealed that while Acr induced oxidative damage and apoptosis, Lut reversed these effects and protected Leydig cells from cellular damage. Also, Lut inhibited the mRNA expressions of Trp53, Casp3, and Bax, which were upregulated by Acr. Consequently, Lut may serve as a potential antioxidant molecule in promoting cell survival by mitigating oxidative damage and apoptosis induced by Acr.