恒清Ⅴ号方对戊四唑诱导癫痫模型小鼠癫痫发作及神经细胞变性损伤的作用机制  

Effect of Hengqing V Formula on Seizures and Degeneration of Nerve Cells in Pentylenetetrazolium Induced Epilepsy Model Mice

作　　者：邓楚珺 孟胜喜[1] 陈慧泽 DENG Chujun;MENG Shengxi;CHEN Huize(Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200233,China)

机构地区：[1]上海交通大学医学院附属第六人民医院,上海200233

出　　处：《中西医结合心脑血管病杂志》2023年第22期4129-4134,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：上海市第六人民医院医疗服务能级提升工程临床医疗技术骨干队伍培育项目(No.20220213);上海市第六人民医院院级课题(No.ynxg202218);上海交通大学医学院“大学生创新训练计划”项目(No.1723Z002)。

摘　　要：目的:探讨恒清Ⅴ号方对戊四唑(PTZ)诱导癫痫模型小鼠癫痫发作及神经细胞变性损伤的作用机制。方法:将无特定病原体(SPF)级雄性C57BL/6J小鼠随机分为正常组、模型组、恒清Ⅴ号方低剂量组、恒清Ⅴ号方中剂量组、恒清Ⅴ号方高剂量组、丙戊酸钠组。给予戊四唑腹腔注射建立癫痫模型。丙戊酸钠组给予丙戊酸钠缓释片溶液灌胃,恒清Ⅴ号方低剂量组、中剂量组、高剂量组分别给予0.267、0.535、1.070 g/mL剂量的恒清Ⅴ号方灌胃,每日2次;正常组和模型组给予等体积生理盐水灌胃,每日2次。各组均持续灌胃30 d。在给药的过程中,除正常组外,其他各组继续腹腔注射戊四唑,每5 d 1次。观察各组在给药后不同时间点癫痫发作级别和发作潜伏期。采用生化法检测小鼠血清及海马组织的超敏C反应蛋白(hs-CRP)、热休克蛋白70水平、大脑皮层和海马中谷氨酸(Glu)含量及谷氨酰胺合成酶(GS)活性;采用Fluoro-Jade B(FJB)染色法分析海马CA1区的神经细胞损伤情况。结果:在给药10 d时,与模型组比较,与清Ⅴ号方中剂量组、高剂量组及丙戊酸钠组小鼠的癫痫发作级别均降低,癫痫发作潜伏期均延长(P<0.05或P<0.01);与恒清Ⅴ号方低剂量组比较,恒清Ⅴ号方中、高剂量组及丙戊酸钠组小鼠的癫痫发作级别均降低,癫痫发作潜伏期均延长(P<0.05或P<0.01);与恒清Ⅴ号方中剂量组、丙戊酸钠组比较,恒清Ⅴ号方高剂量组小鼠的癫痫发作级别均降低,癫痫发作潜伏期均延长(P<0.05)。在给药20、30 d时,与模型组比较,恒清V号组小鼠的癫痫发作级别均降低,癫痫发作潜伏期均延长(P<0.05或P<0.01);与低剂量组比较,恒清Ⅴ号方中剂量组、高剂量组及丙戊酸钠组小鼠的癫痫发作级别均降低,癫痫发作潜伏期均延长(P<0.05或P<0.01);与恒清Ⅴ号方中剂量组、丙戊酸钠组比较,恒清Ⅴ号方高剂量组小鼠的癫痫发作级别均降低,癫Objective:To investigate the effect of Hengqing V Formula(HQ)on seizures and degeneration of nerve cells in pentylenetetrazolium(PTZ)induced epilepsy model mice.Methods:Specific pathogen free(SPF)male C57BL/6J mice were randomly divided into normal group,model group,low dose of HQ(HQ-L)group,medium dose of HQ(HQ-M)group,high dose of HQ(HQ-H)group,and sodium valproate(VPA)group.Except for normal group,the mice in other groups were given pentylenetetrazole(PTZ)intraperitoneal injection to establish epilepsy model.The mice in VPA group were given sodium valproate sustained-release tablet solution by gavage.The mice in HQ-L,HQ-M,and HQ-H groups were given 0.267,0.535,1.070 g/mL of HQ by gavage,both twice a day,respectively.The mice in normal group and model group were given equal volume normal saline by gavage,twice a day.Each group was continuously gavaged for 30 days.In the course of administration,except for the normal group,other groups were to injected with PTZ intraperitoneally once every 5 days.The seizure level and seizure latency at different time points after administration were observed.The levels of high-sensitivity C-reactive protein(hs-CRP)and heat shock protein 70(HSP-70)in serum and hippocampus were measured.The content of glutamate(Glu)and the activity of glutamine synthetase(GS)in cerebral cortex and hippocampus of mice in each group were detected.Fluoro-Jade B(FJB)staining was used to observe the nerve cell injury in hippocampal CA1 area.Results:At the 10th day after administration,compared with model group,the seizure level of mice in HQ-M,HQ-H,and VPA groups decreased and the seizure latency prolonged(P<0.05 or P<0.01).Compared with HQ-L group,the seizure level of HQ-M,HQ-H group,and VPA group decreased,and the seizure latency prolonged(P<0.05 or P<0.01).Compared with HQ-M group and VPA group,the seizure level of HQ-H group decreased and the seize latency was prolonged(P<0.05).At the 20th and 30th day after administration,compared with model group,the seizure grade of the mice in each treatment g

关 键 词：癫痫 恒清Ⅴ号方 谷氨酰胺合成酶活性 超敏C反应蛋白 热休克蛋白 实验研究 

分 类 号：R285.5[医药卫生—中药学]