Study of antioxidant properties
Reactive oxygen species (ROS) are produced during intrinsic cellular metabolism but are also generated by extrinsic agents such as ultraviolet radiations, drugs and other chemicals like environmental pollutants.` ROS induce DNA damage in cells. The accumulation of ROS increases genomic instability. Oxidative stress is thus a factor of mutagenesis, involved in the aging process of cells and carcinogenesis.
The study phosphorylation of histone H2AX (γH2AX) can detect compounds with different genotoxic modes of action, including oxidative stress. Indeed, it has been shown in many studies induction of γH2AX following oxidative stress induced by treatment with (hydrogen peroxide, menadione, KBRO3…).
Thus, it is possible by quantifying the phosphorylation of histone H2AX (INRA license know-how) to determine the antioxidant properties of a compound.
For this study, we mainly work with the HaCat human cell line derived from keratinocytes or the HepG2 hepatic cell line. Cells are treated with an oxidizing product (menadione) in the presence or absence of the compound or mixture to be tested. The N-acetyl-cysteine antioxidant (NAC) is used as a positive control. The γH2AX induction differential makes it possible to demonstrate the antioxidant effect