Inhibition of autophagy-dependent pyroptosis attenuates cerebral ischaemia/reperfusion injury

Autophagy is closely associated with cerebral ischaemia/reperfusion injury, but the underlying mechanisms are unknown. We investigated whether Spautin-1 ameliorates cerebral ischaemia/reperfusion injury by inhibiting Autophagy and whether its derived Pyroptosis is involved in this process. We explored the mechanism of Spautin-1 in cerebral ischaemia/reperfusion. To answer these questions, healthy male Sprague-Dawley rats were exposed to middle cerebral artery occlusion for 60 minutes followed by reperfusion for 24 hours. We found that cerebral ischaemia/reperfusion increased the expression levels of Autophagy and pyroptosis-related proteins. Treatment with Spautin-1 reduced the infarct size and water content and restored some neurological functions. In vitro experiments were performed using oxygen-glucose deprivation/reoxygenation to model PC12 cells. The results showed that PC12 cells showed a significant decrease in cell viability and a significant increase in ROS and Autophagy levels. Spautin-1 treatment reduced Autophagy and ROS accumulation and attenuated NLRP3 inflammasome-dependent Pyroptosis. However, these beneficial effects were greatly blocked by USP13 overexpression, which significantly counteracted the inhibition of Autophagy and NLRP3 inflammasome-dependent Ferroptosis by Spautin-1. Together, these results suggest that Spautin-1 may ameliorate cerebral ischaemia-reperfusion injury via the Autophagy/Pyroptosis pathway. Thus, inhibition of Autophagy may be considered as a promising therapeutic approach for cerebral ischaemia-reperfusion injury.