Anti-oxidative activities and anti-ferroptosis of conditioned medium from umbilical cord mesenchymal stem cells

The therapeutic effect of MSC is closely related to its antioxidant capacity. There is no uniform standard for evaluating the antioxidant capacity of MSC. In this study, we compared the antioxidant capacity of control medium (CON) and conditioned medium (CM) from umbilical cord mesenchymal stem cells cultured for 48 h, about total antioxidant capacity, DPPH scavenging capacity, O2- and hydroxyl radical inhibiting capacity, and the detection of antioxidant enzymes including superoxide dismutase, glutathione peroxidase, and catalase, and resistance to cellular oxidative damage caused by H2O2, SNAP, erastin, and RSL3. The results showed that CM had better DPPH scavenging capacity than CON. No significant differences were observed in antioxidant enzymes. CM did not resist the oxidative damage induced by H2O2 and SNAP, but it had a strong resistance to ferroptosis induced by erastin and RSL3, indicating that CM had excellent resistance to cell lipid peroxidation. CM could improve the cell shrinkage morphology induced by ferroptosis and reduce the production of lipid ROS. qPCR experiments proved that CM improved and regulated multiple pathways of ferroptosis, including genes related to iron metabolism such as FPN, FTH1, TFRC, and IREB2, and redox regulatory genes such as GPX4, AIFM2, DHODH, and TP53, and increased the antioxidant-related transcription factors NRF2 and ATF4.