痫得安丸联合丙戊酸钠通过TNF-α/NF-κB信号通路对癫痫大鼠神经元损伤和炎症的影响*

作者：梁政娆1，娄 勍1，甘海宁2，黄晓丹3

单位：1.肇庆市中医院，广东 肇庆 526020； 2.广东省中医药工程技术研究院，广东 广州 510095； 3.广州中医药大学第五临床医学院，广东 广州 510095

引用：引用：梁政娆，娄勍，甘海宁，黄晓丹.痫得安丸联合丙戊酸钠通过TNF-α/NF-κB信号通路对癫痫大鼠神经元损伤和炎症的影响[J].中医药导报,2025,31(9):24-31.

DOI：10.13862/j.cn43-1446/r.2025.09.004

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摘要：

目的：探究痫得安丸联合丙戊酸钠通过肿瘤坏死因子-α（TNF-α）/核因子κB（NF-κB）信号通路对癫痫模型大鼠神经元损伤和炎症的影响。方法：采用氯化锂-匹罗卡品联合注射复制癫痫大鼠模型，将造模成功的40只癫痫大鼠随机分为模型组（痈+丙联用组）、丙戊酸钠组[0.072 g/（kg·d）]、痫得安丸组[1.350 g/（kg·d）]、痫得安丸[1.350 g/（kg·d）]+丙戊酸钠[0.072 g/（kg·d）]联用组（痫+丙联用组），另取10只健康大鼠作为对照组。给药4周后，记录各组大鼠癫痫发作次数和持续时间，苏木精-伊红（HE）染色及尼氏染色观察海马组织病理学变化，检测血清及海马谷氨酸（Glu）、γ氨基丁酸（GABA）、TNF-α和白细胞介素-1β（IL-1β）水平，RT-qPCR技术检测海马中TNF-α mRNA、肿瘤坏死因子受体1（TNFR1） mRNA、NF-κB p65 mRNA、环氧合酶（COX-2）mRNA、基质金属蛋白酶-9（MMP-9） mRNA表达水平，蛋白质印迹法（Western blotting）检测海马组织TNF-α、TNFR1、NF-κB p65、磷酸化p65（p-p65）、COX-2、MMP-9蛋白表达水平。结果：模型组大鼠癫痫发作次数多于对照组（P＜0.01），癫痫持续时间长于对照组（P＜0.01）；丙戊酸钠组、痫得安丸组、痫+丙联用组大鼠癫痫发作次数少于模型组（P＜0.01），癫痫持续时间均长于模型组（P＜0.01）；痫+丙联用组大鼠癫痫持续时间短于痫得安丸组（P＜0.05）。对照组大鼠海马CA1区锥体神经元排列紧密整齐、结构清晰、染色均匀，胞浆内尼氏小体丰富；模型组大鼠海马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；丙戊酸钠组、痫得安丸组、痫+丙联用组大鼠海马CA1区锥体神经元排列更整齐，染色均匀，尼氏小体数量增多，形态接近对照组大鼠海马。模型组大鼠血清GABA含量低于对照组（P＜0.01），Glu含量、Glu/GABA比值、IL-1β水平、TNF-α水平高于对照组（P＜0.01）；丙戊酸钠组、痫得安丸组及痫+丙联用组大鼠血清Glu含量、Glu/GABA比值、IL-1β水平、TNF-α水平低于模型组（P<0.05或P<0.01）。模型组大鼠大脑GABA含量低于对照组（P<0.05），Glu含量、Glu/GABA比值、IL-1β水平、TNF-α水平高于对照组（P<0.01）；丙戊酸钠组及痫得安丸组大鼠大脑Glu/GABA比值低于模型组（P<0.05）；痫+丙联用组大鼠大脑Glu含量、TNF-α水平、Glu/GABA比值低于模型组（P<0.05或P<0.01）。模型组大鼠海马TNF-α mRNA、TNFR1 mRNA、NF-κB p65 mRNA、COX-2 mRNA、MMP-9 mRNA相对表达量高于对照组（P<0.01）；丙戊酸钠组大鼠海马TNF-α mRNA、MMP-9 mRNA相对表达量低于模型组（P<0.05）；痫得安丸组、痫+丙联用组大鼠海马TNFR1 mRNA、COX-2 mRNA、MMP-9 mRNA、TNF-α mRNA及NF-κB p65 mRNA相对表达量低于模型组（P<0.05或P<0.01）。模型组大鼠海马TNF-α、TNFR1、p-p65、COX-2、MMP-9蛋白相对表达量高于对照组（P<0.05或P<0.01）；丙戊酸钠组大鼠海马TNF-α、TNFR1蛋白相对表达量低于模型组（P<0.05）；痫得安丸组大鼠海马TNF-α、TNFR1、COX-2蛋白相对表达量低于模型组（P<0.05或P<0.01）；痫+丙联用组大鼠海马TNF-α、TNFR1、p-p65、COX-2、MMP-9蛋白相对表达量低于模型组（P<0.01）。结论：痫得安丸与丙戊酸钠合用可减少大鼠癫痫的发生，改善癫痫大鼠神经元损伤，降低炎症反应，其机制可能与抑制TNF-α/NF-κB信号通路有关。

关键词：癫痫；痫得安丸；丙戊酸钠；神经元；炎症；肿瘤坏死因子α；核因子κB；大鼠

Abstract：

Objective: To explore the effects of Xiande'an pill combined with sodium valproate on neuronal injury and inflammation in epileptic rat models through the tumor necrosis factor-α(TNF-α)/nuclear factor-κB (NF-κB) signaling pathway. Methods: Epileptic rat models were established by combined injection of lithium chloride and pilocarpine. Totally 40 successfully modeled epileptic rats were randomly divided into model group, sodium valproate group [0.072 g/(kg·d)], Xiande'an pill group [1.350 g/(kg·d)], and Xiande'an pill [1.350 g/(kg·d)]+sodium valproate [0.072 g/(kg·d)] combination group (combination group). Another 10 healthy rats were used as the control group. After 4 weeks of administration, the frequency and duration of epileptic seizures in each group were recorded. Hematoxylin-eosin staining and Nissl staining were used to observe the pathological changes in the hippocampus. The levels of glutamic acid (Glu), γ aminobutyric acid (GABA), TNF-α and interleukin-1β (L-1β) in serum and brain were detected. The expressions of TNF-α mRNA, tumor necrosis factor receptor 1 (TNFR1) mRNA, NF-κB p65 mRNA, cyclooxygenase 2 (COX-2) mRNA and matrix metalloproteinase 9 (MMP-9) mRNA in the hippocampus were detected by RT-qPCR and Western blotting was used to detect the expressions of TNF-α, TNFR1, NF-κB p65, phosphorylated p65 (p-p65), COX-2 and MMP-9 protein. Results: The model group showed higher frequency of epileptic seizures than control group (P＜0.01), and longer duration of epileptic seizures than  control group (P＜0.01). The  sodium valproate group, Xiande’an pill group and combination group showed lower frequency of epileptic seizures than model group (P＜0.01), and shorter duration of epileptic seizures than model group (P＜0.01). The combination group rats showed shorter duration of epileptic seizures than Xiande’an pill group (P＜0.01). The pyramidal neurons in the hippocampal CA1 region of the control group rats were arranged tightly and neatly, with clear structure and uniform staining, and abundant Nissl bodies in the cytoplasm. The pyramidal neurons in the hippocampal CA1 area were disordered in arrangement, and the cells atrophied in model group. The staining deepened in model group. The number of Nisslakosomes was significantly reduced. The pyramidal neurons in the CA1 region of the hippocampus are arranged more neatly and stained uniformly in the sodium valproate group, Xiande'an pill group and combination group, with an increase in the number of Nissl bodies and a morphology similar to the control group. The model group showed lower serum GABA content than control group (P＜0.01), while higher serum Glu, Glu/GABA, IL-1β and TNF-α than control group (P＜0.01). Sodium valproate group, Xiande'an pill group and combination group showed lower serum Glu, Glu/GABA, IL-1β and TNF-α than model group (P＜0.05 or P＜0.01). The model group showed lower GABA content in brain than control group  (P＜0.01), while higher serum Glu, Glu/GABA, IL-1β and TNF-α in brain than control group (P＜0.01). Sodium valproate group and Xiande'an pill group showed lower brain Glu/GABA than model group (P＜0.05). Combination group showed lower Glu, TNF-α and Glu/GABA in brain, than model group (P＜0.05 or P＜0.01). The model group showed higher relative expression levels of TNF-α mRNA, TNFR1 mRNA, NF-κB p65 mRNA, COX-2 mRNA and MMP-9 mRNA than control group (P<0.01). Sodium valproate group showed lower relative expression levels of TNF-α mRNA and MMP-9 mRNA than model group (P<0.05). Xiande'an pill group and combination group showed lower TNFR1 mRNA, COX-2 mRNA, MMP-9 mRNA, TNF-α mRNA and NF-κB p65 mRNA than model group (P＜0.05 or P＜0.01). The model group showed higher relative expression levels of TNF-α, TNFR1, p-p65, COX-2 and MMP-9 protein than control group (P＜0.05 or P＜0.01). Sodium valproate group showed lower relative expression levels of TNF-α and TNFR1 protein than model group (P<0.05). Xiande'an pill group  showed lower TNF-α, TNFR1 and COX-2 protein than model group (P＜0.05 or P＜0.01). Combination group showed lower TNF-α, TNFR1, p-p65, COX-2 and MMP-9 protein than model group (P＜0.01). Conclusion: Xiande'an pill combined with sodium valproate can reduce the occurrence of epilepsy in rats, improve pathological injury, protect neurons and reduce inflammatory response of epileptic rats, the mechanism of which is related to inhibiting TNF-α/NF-κB signaling pathway.

Key words：epilepsy; Xiande'an pill; sodium valproate; neurons; inflammation; TNF-α; NF-κB; rat

发布时间：2026-01-08

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