Porcine Cardiac Blood processed Kansui Radix alleviates PTZ-induced epileptic damage in mice via the bidirectional regulation of GABRA1 and cGMP/PKG signaling pathway.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: According to Menghe Medical School, Porcine Cardiac Blood (PCB) enhances the upward distribution of medicinal substances throughout the body. Although both ancient and modern evidence suggest that PCB processed Kansui Radix (PCB-GS) has antiepileptic effects, its mechanisms for epilepsy remain unclear. AIM OF THE STUDY: To assess the therapeutic effects and mechanisms of PCB-GS in pentylenetetrazol (PTZ)-induced epileptic damage in mice. MATERIALS AND METHODS: Quantitative analysis of chemical profiles of PCB-GS and Kansui Radix (GS) extracts was performed using UPLC-Q-TOF-MS/MS technology. Characterization of extracts was conducted via scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Ultraviolet spectrum. An acute epileptic model in mice induced by PTZ was established. Hippocampal proteomics and molecular biology techniques were employed to investigate the mechanisms by which PCB-GS treats epilepsy. RESULTS: Following PCB processing, the liposoluble components in GS increased, forming globular-like particles and altering UV spectral characteristics. PCB-GS alleviated PTZ-induced epileptic seizures, including reduced seizure frequency and severity, prolonged seizure latency, shortened total seizure duration, and suppression of spike counts. PCB-GS alleviated neuronal damage in the hippocampal dentate gyrus (DG), reduced the expression of the epilepsy marker c-Fos, and decreased microglial activation. Hippocampal proteomics identified the cGMP/PKG signaling pathway as a key mediator of PCB-GS effects. Western blot analysis showed that cGMP, p-PKG and p-VASP, levels increased significantly 1h after PTZ administration, then decreased markedly by 24h. PCB-GS modulates the cGMP/PKG signaling pathway by bidirectionally regulating the expression of the upstream GABRA1 protein, ultimately suppressing the elevation of downstream p-CREB and Cleaved caspase-3 protein levels. CONCLUSION: PCB-GS protected against epileptic seizures via bidirectional modulation of the cGMP/PKG signaling axis through stabilization of GABRA1 expression, thereby suppressing p-CREB and Cleaved caspase-3.