Ozone treatment attenuates the ferroptosis induced by cerebral ischemia/reperfusion injury via activation of the Nrf2/Slc7a11/Gpx4 signaling pathway

Abstract

Abstract Background The pathogenesis of brain ischemic/reperfusion (I/R) insult is characterised by the loss of neurons as a result of excessive oxidative stress responses. A form of oxidative cell death known as ferroptosis can be triggered when there is a breakdown in the equilibrium that exists between antioxidants and pro-oxidants in cells. As a natural bioactive molecule with antioxidant/anti-apoptotic and pro-autophagic properties, ozone can enhance the capacity of the antioxidant system and ameliorate oxidative stress. Yet, the mechanism of its role in neuronal ferroptosis remains unclear. Therefore, we investigated the functions and possible mechanisms of ozone in cerebral I/R-induced ferroptotic neuronal death. Methods A model of cerebral ischemia-reperfusion injury was created in S-D rats that had been pretreated with ozone. Intraperitoneal administration of the Nrf2 inhibitor ML385, the Slc7a11 inhibitor Erastin, and the Gpx4 inhibitor RSL3 was performed 1h prior to the creation of the model. Results According to the findings of our research, ozone preconditioning was able to mitigate neuronal damage caused by cerebral ischemia-reperfusion (I/R), lessen the severity of neurological deficits, lower the volume of cerebral infarcts, and reduce cerebral infarct volume in MCAO rats. One possible mechanism for this protective effect is the suppression of neuronal ferroptosis. Transmission electron microscopy, immunofluorescence, and Western blotting findings all pointed to ferroptosis in the aftermath of MCAO-induced brain damage. The present study found that MCAO caused morphological damage to neuronal mitochondria, enhanced the accumulation of lipid peroxidation, and promoted MDA production. Moreover, MCAO decreased the levels of FTH1 and GPX4, which act as negative regulators of ferroptosis, and increased the levels of ACSL4, which acts as a positive regulator of ferroptosis. Ozone preconditioning has been shown to have a protective impact on neuronal by increasing the nuclear translocation of Nrf2 and the expression of Slc7a11 and Gpx4. Meanwhile, treatment with ML385, Erastin and RSL3 significantly reversed the protective effect of ozone preconditioning on neuronal ferroptosis. Conclusion Ozone treatment attenuates the ferroptosis in cerebral ischemia/reperfusion injury rat model via Nrf2/Slc7a11/Gpx4 Pathway, which lays a new theoretical foundation for the use of ozone as a possible therapy to prevent ischemic stroke.