Hesperetin alleviates neuronal pyroptosis by promoting mitophagy via DHCR24/BACE1 signaling pathway after subarachnoid hemorrhage in mice.

Pyroptosis is a new type of programmed cell death that plays an important role in neuronal injury after subarachnoid hemorrhage (SAH). The effect of natural compounds in SAH has attracted much attention. Hsperetin has been found to have neuroprotective effects in ischemic stroke, but its role in SAH has not been studied. An in vivo model of SAH in male C57BL/6 mice is constructed by the endovascular perforation method, and the heme intervention HT22 cells simulate the in vitro SAH model. After administration of hesperetin, SAH grade, brain water content (BWC), modified garcia score, neurobehavior tests, cerebral blood flow, transmission electron microscope, western blot, and immunofluorescence staining were conducted. Our study found that hesperetin significantly alleviated neurobehavioral deficits, improved cerebral blood flow, and reduced the expression of NLRP3, GSDMD-N, Caspase-1, ASC, IL-18, which were also demonstrated in vitro. Mechanistically, hesperetin notably promoted mitophagy by regulating DHCR24 and BACE1, thereby inhibiting neuronal pyroptosis. This effect was eliminated by U18666A and 3-MA administration. Our findings demonstrated that hesperetin alleviated neuronal pyroptosis by promoting mitophagy via DHCR24/BACE1 to provide neuroprotective effects on EBI. These results suggest that hesperetin has the potential to be a therapeutic target for SAH.