中国现代神经疾病杂志 ›› 2025, Vol. 25 ›› Issue (4): 303-310. doi: 10.3969/j.issn.1672-6731.2025.04.007

• 基础研究 • 上一篇    下一篇

2 甘草酸对海人藻酸癫痫大鼠模型癫痫发作及海马神经元损伤的影响

陈艳, 余艳华, 陈季南, 李圣华, 李军荣*()   

  1. 211100 南京医科大学附属江宁医院神经内科
  • 收稿日期:2025-02-25 出版日期:2025-04-25 发布日期:2025-05-19
  • 通讯作者: 李军荣
  • 基金资助:
    江苏省南京市卫生和计划生育委员会医学科技发展项目(YKK17221)

Effect of glycyrrhizin acid on the seizure onset and hippocampal neuron injury in kainic acid induced epileptic rat models

Yan CHEN, Yan-hua YU, Ji-nan CHEN, Sheng-hua LI, Jun-rong LI*()   

  1. Department of Neurology, The Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing 211100, Jiangsu, China
  • Received:2025-02-25 Online:2025-04-25 Published:2025-05-19
  • Contact: Jun-rong LI
  • Supported by:
    Medical Science and Technology Development Project of Nanjing Health and Family Planning Commission in Jiangsu(YKK17221)

摘要:

目的: 探讨甘草酸对海人藻酸癫痫大鼠癫痫发作和海马神经元损伤的影响及可能作用机制。方法: 于大鼠海马组织注射海人藻酸制备癫痫模型,随机分为假手术组、癫痫模型组和甘草酸干预组(25 mg/kg组和50 mg/kg组),观察各组癫痫发作潜伏期及严重程度、监测急性发作期异常脑电活动,免疫组化染色和免疫印迹法检测海马CA3区神经元损伤程度及高迁移率族蛋白1(HMGB1)表达变化。结果: 与模型组相比,甘草酸25 mg/kg组和50 mg/kg组大鼠癫痫发作潜伏期延长[(109.33±42.84)min对(51.17±22.31)min,t=-2.950,P=0.015;(109.50±35.79)min对(51.17±22.31)min,t=-3.388,P=0.007],模型制备后6 h内达≥ Ⅳ级发作频次减少[(2.83±0.75)次对(5.00±1.55)次,t=3.081,P=0.012;(2.67±1.75)次对(5.00±1.55)次,t=2.445,P=0.035],脑电图痫样放电减少。癫痫发作急性期,模型组大鼠海马CA3区神经元数目减少且少于假手术组[(40.33±5.69)个对(72.33±7.51)个;t=5.886,P=0.004],甘草酸25 mg/kg组和50 mg/kg组神经元数目增加且多于模型组[(58.33±2.52)个对(40.33±5.69)个,t=-5.014,P=0.007;(57.00±6.25)个对(40.33±5.69)个,t=-3.418,P=0.027]。免疫组化染色显示,模型组大鼠海马CA3区HMGB1表达水平[积分光密度值(IOD)]升高且高于假手术组[(3.79±0.50)×106 IOD对(2.16±0.45)×106 IOD;t=-4.216,P=0.014],甘草酸25 mg/kg组和50 mg/kg组海马CA3区HMGB1表达水平下降且低于模型组[(2.50±0.52)×106 IOD对(3.79±0.50)×106 IOD,t=3.090,P=0.037;(2.66±0.44)×106 IOD对(3.79±0.50)×106 IOD,t=2.955,P=0.042]。免疫印迹法显示,模型组大鼠海马组织HMGB1表达水平(相对灰度值)高于假手术组(1.19±0.17对0.54±0.14;t=-5.078,P=0.007),甘草酸25 mg/kg组和50 mg/kg组海马组织HMGB1表达降低且低于模型组(0.65±0.04对1.19±0.17,t=5.286,P=0.028;0.58±0.13对1.19±0.17,t=4.953,P=0.008)。结论: 甘草酸可延长癫痫模型大鼠发作潜伏期、降低发作严重程度、减少脑电图痫样放电,并缓解海马神经元损伤,其作用机制可能与抑制海马组织HMGB1表达有关。

关键词: 癫痫, 红藻氨酸, 甘草酸, 海马, 神经元, 高迁移率族蛋白质类, 免疫组织化学, 印迹法,蛋白质, 大鼠, 疾病模型,动物

Abstract:

Objective: To explore the effect and the possible mechanism of glycyrrhizin acid (GA) on the seizure onset and hippocampal neuron injury in kainic acid (KA) induced epileptic rat models. Methods: The epilepsy model was established by injecting KA into the rat hippocampus. Rats were randomly divided into sham group, epilepsy group (EP group), and GA group (25 mg/kg group and 50 mg/kg group). After injection, the latency period and severity of seizure of rats in each group were observed. The acute phase EEG of rats in different groups were monitored. The neuron injury in hippocampus CA3 region and the expression of high-mobility group box 1 (HMGB1) of rats in different groups were detected by immunohistochemistry staining and Western blotting. Results: Compared to the EP group, the epileptic seizure latency of GA 25 mg/kg group and 50 mg/kg group significantly lengthened [(109.33±42.84) min vs. (51.17±22.31) min, t =-2.950, P = 0.015; (109.50±35.79) min vs. (51.17±22.31) min, t =-3.388, P = 0.007], the times of seizure ≥ Ⅳ grade during 6 h after injection in GA 25 mg/kg group and 50 mg/kg group significantly reduced [(2.83±0.75) times vs. (5.00±1.55) times, t = 3.081, P = 0.012; (2.67±1.75) times vs. (5.00±1.55) times, t = 2.445, P = 0.035]. After the treatment of GA, the number of spike and sharp waves in the acute phase EEG obviously decreased. In the acute stage of epileptic seizure, the neurons number in hippocampus CA3 region of EP group reduced, and was significantly less than that of sham group [(40.33±5.69) neurons vs. (72.33±7.51) neurons, t = 5.886, P = 0.004]; while the number of neurons in hippocampus CA3 region of GA 25 mg/kg group and 50 mg/kg group increased, and was significantly more than that of EP group [(58.33±2.52) neurons vs. (40.33±5.69) neurons, t =-5.014, P = 0.007; (57.00±6.25) neurons vs. (40.33±5.69) neurons, t =-3.418, P = 0.027]. The expression of HMGB1 [integrated optical density (IOD)] in hippocampus CA3 region of EP group heightened, and was significantly higher than that of sham group [(3.79±0.50)×106 IOD vs. (2.16±0.45)×106 IOD, t =-4.216, P = 0.014]; while the expression of HMGB1 in hippocampus CA3 region of GA 25 mg/kg group and 50 mg/kg group decreased, and was significantly lower than that of EP group [(2.50±0.52)×106 IOD vs. (3.79±0.50)×106 IOD, t = 3.090, P = 0.037; (2.66±0.44)×106 IOD vs. (3.79±0.50)×106 IOD, t = 2.955, P = 0.042]. The expression of HMGB1 protein (relative gray value) in hippocampus tissue detected of EP group increased, and was significantly higher than that of sham group (1.19±0.17 vs. 0.54±0.14, t =-5.078, P = 0.007); while the expression of HMGB1 protein in hippocampus tissue of GA 25 mg/kg group and 50 mg/kg group lessened, and was significantly lower than that of EP group (0.65±0.04 vs. 1.19±0.17, t = 5.286, P = 0.028; 0.58±0.13 vs. 1.19±0.17, t = 4.953, P = 0.008). Conclusions: GA can prolong the epileptic seizure latency, reduce the seizure severity and epileptic discharges, and alleviate the hippocampal neuron injury in KA induced epileptic rats. These effects may be associated with its inhibition on the expression of HMGB1.

Key words: Epilepsy, Kainic acid, Glycyrrhizic acid, Hippocampus, Neurons, High mobility group proteins, Immunohistochemistry, Blotting, Western, Rats, Disease models, animal