Purification and characterization of antioxidant peptides from cooked eggs using a dynamic in vitro gastrointestinal model in vascular smooth muscle A7r5 cells

Antioxidant effect of fractions from defatted digests on A7r5 cells. Untr: control; F1, F2, and F3: eluted from a C18 cartridge column with 20, 50, and 100% of ACN solution containing 0.1% TFA, subsequently; whole digest: the original digest was simply defatted and desalted. Cultured monolayers of vascular smooth muscle cells (A7r5 cell line) were pretreated with various fractions (with concentration of 2.5？mg/mL) for 2？h and without pretreatment as control. The cells were then washed once and incubated at 37？°C with 10？μM DHE in the DMEM containing 1% FBS for 30？min. Fluorescence intensity from three randomly chosen fields was measured by its mean fluorescence intensity per cell (MFI/cell). Superoxide generation was calculated as fold change in MFI/cell and presented as percentage (%) of the untreated control. Mean？±？SEM of three independent experiments are shown. Asterisk indicates p？<？0.05 compared to the untreated contro