Effects of astaxanthin on oxidative stress response in primary mouse hepatocytes

Objective  To investigate the effect of astaxanthin on H₂O₂-induced oxidative stress response in primary mouse hepatocytes.

Method  Primary mouse hepatocytes were isolated by an in situ two-step perfusion technique. The model of oxidative stress response in primary mouse hepatocytes was established using H₂O₂ treatment and malondialdehyde(MDA) as an indicating index. The content of reative oxygen species (ROS) and change in apoptosis rate were measured by flow cytometry. The contents of MDA and glutathione(GSH), and the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) were measured by biochemical methods. The relative mRNA expression levels of antioxidant enzymes including SOD and GSH-px were measured by fluorescence quantitative PCR. The relative expression of Nrf2 protein was measured by Western-blot.

Result  The model of oxidative stress response performed the best with the primary mouse hepatocytes pre-protected by 5 μg·mL^–1 astaxanthin for 3 h before being exposed to 10 μmol·L^–1H₂O₂ for 3 h. Astaxanthin decreased the apoptosis rate and the contents of ROS, MDA and GSH in primary mouse hepatocytes induced by H₂O₂. Astaxanthin increased the activities of SOD and GSH-px as well as the relative mRNA expression levels of SOD and GSH-px, and inhibited the nuclear transfer of Nrf2 protein.

Conclusion  Astaxanthin has protective effects on H₂O₂-induced primary mouse hepatocytes with oxidative stress.