Neuroprotective effect of glycosides in Buyang Huanwu Decoction on pyroptosis following cerebral ischemia-reperfusion injury in rats

Ethnopharmacological relevance: Buyang Huanwu Decoction (BYHWD) is used in classical traditional Chinese medicine to prevent and treat cerebral ischemia. Glycosides, which are effective components extracted from BYHWD, mainly include astragaloside IV, paeoniflorin, and amygdalin. These glycosides are the primary pharmacologically effective constituents of BYHWD that act against cerebral ischemic nerve injury; however, the mechanism of action of BYHWD is still unclear. Aim of the study: The present study aimed to determine the effect of BYHWD glycosides on pyroptosis after cerebral ischemia reperfusion injury and explore whether its mechanism involves the classical pyroptosis pathway mediated by NLRP3. Material and methods: Adult male Sprague-Dawley rats (n = 140) were randomly divided into seven groups: sham, cerebral ischemia and reperfusion (I/R), glycosides (0.064 g/kg, 0.128 g/kg, and 0.256 g/kg), BYHWD, and AC-YVAD-CMK (caspase-1 inhibitor). A rat model of cerebral I/R was established via classic middle cerebral artery occlusion (MCAO) for 2 h, followed by 24-h reperfusion. Neurological function was estimated using neurological defect scores. Brain infarct volumes were determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and nerve cell damage was evaluated by Nissl staining. Pyroptosis was detected using TUNEL and caspase-1 immunofluorescence double staining. Protein expression of NLRP3, ASC, caspase-1, pro-caspase-1, and IL-1 beta was analyzed using Western blot analysis. Results: Glycosides improved neurological dysfunction, alleviated neuronal damage, and inhibited neuronal pyroptosis. The 0.128 g/kg glycosides group showed the most significant effects. Furthermore, we observed that this group showed significant inhibition of the expression of NLRP3, ASC, pro-caspase-1, caspase-1, and IL-1 beta proteins of the NLRP3-mediated classical pathway of pyroptosis. Conclusions: Glycosides exert neuroprotective effects by inhibiting pyroptosis of neurons after cerebral I/R injury. The underlying mechanism of action is closely related to the regulation of the classical pyroptosis pathway by NLRP3.