*4.9. Lipid Peroxidation Measurement*

Lipid peroxidation was measured using the Lipid Peroxidation (MDA) Assay Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's instructions. Extensive oxidative species generation can lead to the formation of malondialdehyde (MDA) as a result of the peroxidation of polyunsaturated fatty acids (PUFAs), which contain at least three double bonds. MDA can react with thiobarbituric acid (TBA) to form a colorimetric, as well a fluorescent, product, which is a well-known biomarker of the peroxidation of polyunsaturated lipids [65]. The tissues were lysed on ice using MDA lysis buffer containing 3 μL of butylated hydroxytoluene (BHT), and the samples were centrifuged at 13,000× *g* for 10 min. Next, a TBA solution was added and incubated at 95 ◦C for 60 min. The samples were chilled to room temperature in an ice bath for 10 min and transferred to a black bottomed 96-well plate. Optical density was measured at 532 nm using an ELISA plate reader.
