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.