Abstract：
It is widely acknowledged that epilepsy is a neurological disorder characterized by recurrent and atypical neuronal discharges, resulting in transient dysfunction within the brain. The protective role of hydrogen sulfide (H2S) in epilepsy has been elucidated by recent studies, but the underlying mechanisms remain poorly understood. To investigate this, the concentration of H2S was measured by spectrophotometry and a fluorescent probe in LiCl/Pilocarpine (LiCl/Pilo)-induced seizures in rats. The localization of proteins was examined using immunofluorescence. Electroencephalogram and behavioral tests were employed to evaluate the occurrence of seizures. Neuropathological changes in the hippocampus were examined by hematoxylin-eosin staining, Nissl staining, and transmission electron microscopy. Through proteomics and bioinformatics analysis, we identified the differential proteins in the hippocampus of rats following H2S intervention. Protein changes were detected through western blotting. The results showed that H2S treatment significantly alleviated seizures and minimized post-seizures neurological damage in rats. Proteomics analysis revealed adenylate cyclase 3 (AC3) as a protein potentially targeted by H2S. Moreover, the AC3 activator forskolin reversed the downregulation effect of H2S on the AC3/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/transient receptor potential vanilloid 2 (TRPV2) signaling pathway. In conclusion, H2S targets and downregulates the expression of AC3, thereby modulating the AC3/cAMP/PKA signaling pathway to regulate the expression of TRPV2 in LiCl/Pilo-induced seizures, ultimately leading to seizure inhibition and neuroprotection.
摘要：
人们普遍认为癫痫是一种神经系统疾病，其特征是复发性和非典型的神经元放电，导致大脑内的短暂功能障碍。最近的研究已经阐明了硫化氢（H2S）在癫痫中的保护作用，但潜在的机制仍然知之甚少。为了调查这一点，通过分光光度法和荧光探针测量LiCl/毛果芸香碱（LiCl/Pilo）诱导的大鼠癫痫发作中H2S的浓度。使用免疫荧光检查蛋白质的定位。采用脑电图和行为测试来评估癫痫发作的发生。通过苏木精-伊红染色检查海马的神经病理变化，尼氏染色,和透射电子显微镜。通过蛋白质组学和生物信息学分析,我们鉴定了H2S干预后大鼠海马中的差异蛋白。通过蛋白质印迹检测蛋白质变化。结果表明，H2S处理可明显减轻大鼠的癫痫发作，并最大程度地减少癫痫发作后的神经损伤。蛋白质组学分析显示腺苷酸环化酶3（AC3）是H2S潜在靶向的蛋白质。此外，AC3激活剂Forskolin逆转了H2S对AC3/环磷酸腺苷（cAMP）/蛋白激酶A（PKA）/瞬时受体电位香草酸2（TRPV2）信号通路的下调作用。总之,H2S靶向下调AC3的表达,从而调节AC3/cAMP/PKA信号通路,从而调节TRPV2在LiCl/Pilo诱导的癫痫发作中的表达,最终导致癫痫抑制和神经保护。
