Tongfu-Xingshen Capsule ameliorates ischemic stroke by inhibiting pyroptosis-mediated neuroinflammation.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Ischemic stroke (IS) represents a highly prevalent acute cerebrovascular disorder in clinical practice, featured by ischemic-hypoxic brain damage secondary to inadequate cerebral perfusion. This devastating condition is associated with substantial disability and recurrence rates, imposing a severe burden on patients' survival and quality of life. Tongfu-Xingshen Capsule (TXC), a conventional Chinese medicinal agent clinically indicated for intracerebral hemorrhage and IS management, exerts favorable effects in mitigating stroke-associated pneumonia and preserving the structural integrity of the blood-brain barrier. Nevertheless, the precise molecular mechanisms through which TXC exerts its therapeutic benefits in IS warrant further investigation. AIM OF THE STUDY: This study aimed to clarify the therapeutic efficacy and underlying mechanisms of TXC against IS, with a particular focus on regulating microglia-mediated neuroinflammatory responses. MATERIALS AND METHODS: Middle cerebral artery occlusion (MCAO) model was established in rats for in vivo experiments, while the oxygen-glucose deprivation/reperfusion (OGD/R) model was constructed in BV2 microglia for in vitro studies. Methods including TTC staining, commercial kits, RT-qPCR, Western blot, and immunofluorescence were used to evaluate the effects of TXC on cerebral infarct volume, oxidative stress, microglial polarization, and changes in the RhoA/ASC/GSDMD signaling pathway. RESULTS: TXC reduced the cerebral infarct volume in MCAO rats, decreased the level of oxidative stress in the brain, and ameliorated neurological deficits. Mechanistic studies revealed that the expression of RhoA in microglia is decreased after ischemia-reperfusion injury, and TXC upregulated RhoA, inhibited Rock2/ASC-mediated pyroptosis, and regulated microglial activation. Further in vitro experiments demonstrated that TXC promoted the restoration of M1/M2 polarization balance in OGD/R-induced BV2 cells, while also upregulating RhoA and suppressing Rock2/ASC-mediated pyroptosis. However, the positive effects of TXC on OGD/R-induced BV2 cells were abrogated following intervention with a RhoA inhibitor. CONCLUSIONS: TXC can inhibit microglia-mediated neuroinflammatory injury and oxidative stress by regulating the RhoA/ASC/GSDMD signaling pathway, thereby alleviate IS-induced cerebral injury.