【目的】 通过体内外研究，探讨痫得安丸联用丙戊酸钠对卒中后癫痫的改善作用及机制。【方法】（1）动物实验：采用 单侧大脑中动脉闭塞结合结扎颈总动脉（MCA/CCAo）法复制大鼠缺血性卒中后癫痫模型。将造模成功的40只癫痫大鼠随机分 为模型组、丙戊酸钠组、痫得安丸组、痫+丙联用组，另取10只大鼠仅暴露血管后即原路缝合切口作为假手术组。给药4周 后，记录各组大鼠癫痫发作频率和持续时间，苏木精-伊红（HE）染色法观察海马组织病理学变化，尼氏染色法观察海马组织 尼氏小体情况，检测大脑谷氨酸（Glu）、γ氨基丁酸（GABA）、肿瘤坏死因子α（TNF-α）和白细胞介素1β（IL-1β）水平，实时 定量聚合酶链反应（RT-qPCR）、Western Blot 法分别检测海马组织 TNF-α、肿瘤坏死因子受体 1（TNFR1）、核转录因子 κB （NF-κB）p65、磷酸化 p65（p-p65）、环氧化酶（Cox2）、基质金属蛋白酶 9（MMP9）、原癌基因 Fos（c-Fos）的 mRNA、蛋白表 达。（2）细胞实验：将RAW264.7细胞接种于96孔板，分为空白组、脂多糖（LPS）组、丙戊酸钠组、痫得安丸组、痫+丙联用组，孵育 6 h 后，加入 LPS 培养 24 h。RT-qPCR、Western Blot 法分别检测 RAW264.7 细胞中 IL-1β、TNF- α、TNFR1、 NF-κB p65、p-p65、Cox2、MMP9 的mRNA、蛋白表达。【结果】（1）动物实验：与假手术组比较，模型组大鼠的癫痫发作 次数和持续时间显著升高（P＜0.01），海马CA1区的锥体神经元排列紊乱、细胞萎缩、染色加深，尼氏小体数量明显减少，大 鼠大脑 GABA 含量降低，Glu 含量，Glu/GABA 比值，IL-1β、TNF-α 水平升高（P＜0.01），海马 TNF-α、TNFR1、NF-κB p65、Cox2、MMP9、c-Fos 基因表达升高（P＜0.01），TNF-α、TNFR1、p-p65、Cox2、MMP9 蛋白表达升高（P＜0.05 或 P＜ 0.01）。痫+丙联用组大鼠癫痫发作频率及持续时间显著降低，海马病理观察可见神经元排列规整性、胞体轮廓清晰度及核质 染色均匀性及尼氏小体数量分布情况明显改善，大脑组织GABA含量升高，Glu含量，Glu/GABA比值，IL-1β、TNF-α水平 降低，海马 TNF-α、TNFR1、NF-κB p65、Cox2、MMP9、c-Fos 基因表达水平升高，海马 TNF-α、TNFR1、p-p65、Cox2、 MMP9蛋白表达水平降低，与模型组比较，差异均有统计学意义（P＜0.05或P＜0.01），且治疗效果优于丙戊酸钠组或痫得安 丸组（P＜0.05 或 P＜0.01）。（2）细胞实验：LPS 刺激后各给药组 RAW264.7 细胞 IL-1β、Cox2、MMP9 基因表达及 TNF-α、pp65、Cox2、MMP9蛋白表达均较模型组下降（P＜0.05或P＜0.01），丙戊酸钠组、痫+丙联用组RAW264.7细胞TNF-α基因表 达较模型组下降（P＜0.05），TNFR1蛋白表达较模型组下降（P＜0.01），痫+丙联用组细胞MMP9基因及p-p65蛋白表达较丙 戊酸钠组下降（P＜0.01），痫+丙联用组细胞TNF-α、TNFR1蛋白表达较痫得安丸组下降（P＜0.05）。【结论】 痫得安丸与丙 戊酸钠合用可改善卒中后癫痫大鼠癫痫的发生和神经炎症反应，其机制可能与抑制TNF-α/NF-κB信号通路有关。

Objective To investigate the therapeutic effect and mechanism of Xiande’an Pills （XAP） combined with sodium valproate （VPA） on post-stroke epilepsy （PSE） through in vivo and in vitro studeies. Methods （1） Animal experiment：A post-ischemic stroke epilepsy model was established in rats using unilateral middle cerebral artery occlusion combined with common carotid artery ligation （MCA/CCAo）. Forty successfully modeled epileptic rats were randomly divided into the model group，VPA group，XAP group，and XAP+VPA combination group. Ten additional rats underwent sham surgery （vessel exposure only followed by wound closure） as the sham group. After 4 weeks of administration，seizure frequency and duration were recorded. Hippocampal histopathology was observed by hematoxylin-eosin （HE） staining，Nissl bodies were assessed by Nissl staining，and levels of glutamate （Glu），γ-aminobutyric acid （GABA），tumor necrosis factor-α（TNF-α），and interleukin-1β（IL-1β） in the brain were measured. mRNA and protein expression of TNF- α， TNF receptor 1（TNFR1）， nuclear factor- κB （NF-κB） p65，phosphorylated p65（p-p65），cyclooxygenase-2（Cox2），matrix metalloproteinase-9 （MMP9），and c-Fos in hippocampal tissue were detected by RT-qPCR and Western Blot.（2） Cell experiment： RAW264.7 cells were seeded in 96-well plates and divided into blank control，lipopolysaccharide （LPS），VPA， XAP，and XAP+VPA combination groups. After 6 hours of incubation，LPS was added for 24 hours. mRNA and protein expression of IL-1β，TNF-α，TNFR1，NF-κB p65，p-p65，Cox2，and MMP9 in RAW264.7 cells were detected by RT-qPCR and Western Blot. Results（1） Animal experiment：Compared with the sham group， the model group showed significantly increased seizure frequency and duration （P＜0.01），disorganized pyramidal neuron arrangement，cell atrophy，and deepened staining in the hippocampal CA1 region，significantly reduced Nissl body count，decreased GABA content in serum and brain，increased Glu content，Glu/GABA ratio，and IL-1β and TNF-α levels （P＜0.01），upregulated hippocampal mRNA expression of TNF-α，TNFR1，NF-κB p65，Cox2，MMP9 and c-Fos （P＜0.01），and increased protein expression of TNF-α，TNFR1，p-p65，Cox2 and MMP9（P＜0.05 or P＜0.01）. The XAP+VPA groups exhibited significantly reduced seizure frequency and duration， demonstrated superior improvements in neuronal arrangement regularity， cell body contour clarity， uniform nuclear staining，and Nissl body distribution，increased GABA content brain，decreased Glu content， Glu/GABA ratio，and IL-1β and TNF-α levels，downregulated hippocampal mRNA expression levels of TNF-α， TNFR1，NF-κB p65，Cox2，MMP9 and c-Fos，and decreased protein expression levelsof TNF-α，TNFR1， p-p65，Cox2 and MMP9，the differences being statistically significant compared with the model group （P＜0.05 or P＜0.01），and the curative effects were superior to the XAP group or VPA group （P＜0.05 or P＜0.01）.（2）Cell experiment：After LPS stimulation，all treatment groups showed decreased mRNA expression of IL-1β，Cox2， and MMP9，and decreased protein expression of TNF-α，p-p65，Cox2，and MMP9 compared to the LPS group （P＜0.05 or P＜0.01）. The VPA and XAP+VPA groups exhibited decreased TNF-α mRNA expression （P＜0.05） and TNFR1 protein expression （P＜0.01） compared to the LPS group. The XAP+VPA group showed lower MMP9 mRNA and p-p65 protein expression than the VPA group （P＜0.01）， and lower TNF- α and TNFR1 protein expression than the XAP group （P＜0.05）. Conclusion The combination of XAP and VPA alleviates seizures， neuroinflammatory responses in rats with post-stroke epilepsy，potentially through inhibition of the TNF-α/NF-κB signaling pathway.

R285.5

广东省中医药局科研基金重点项目（编号：20223024）；国家中医药管理局第五批全国中医临床优秀人才研修项目（国中医药人教函 〔2022〕 1 号）