人参皂苷Rb_(1)治疗性给药的抗癫痫作用及机制研究  

Study on the antiepileptic effect and mechanism of ginsenoside Rb_(1)

作　　者：陈雪 张慧明 马青[1] 徐梦 李娟 柳钰书 刘滢 王茜[1] 唐民科[1] CHEN Xue;ZHANG Huiming;MA Qing;XU Meng;LI Juan;LIU Yushu;LIU Ying;WANG Xi;TANG Minke(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《西北药学杂志》2021年第5期739-745,共7页Northwest Pharmaceutical Journal

基　　金：中南中医药防治重大疾病国际合作基地项目(国家中医局国际化专项,编号0610-2040N F020928);教育部专项经费(基本科研业务费)(编号:1000061223731);“一带一路”中医药发展与中药学科建设关键技术研究(2020科技部高端外国专家引进计划,编号:DL 20200001008)。

摘　　要：目的探讨人参皂苷Rb_(1)治疗性给药的抗癫痫作用及其作用机制。方法采用戊四唑(PTZ)点燃的方法建立小鼠癫痫模型。将造模成功的小鼠随机分为PTZ组、丙戊酸钠(VPA)组和人参皂苷Rb_(1)低(10 mg·kg^(-1))、中(20 mg·kg^(-1))、高(40 mg·kg^(-1))剂量组。各给药组分别腹腔注射相应药物30 d,每日1次。通过观察给药后不同时间点癫痫发作级别和发作潜伏期,评估小鼠癫痫发作的严重程度和治疗性给药的恢复情况。采用生化法检测各组小鼠大脑皮层和海马中谷氨酸(Glu)含量和谷氨酰胺合成酶(GS)活性;Fluoro-Jade B(FJB)染色观察海马CA1区的神经细胞损伤情况;逆转录-聚合酶链反应(RT-PCR)法检测小鼠大脑皮层和海马中谷氨酸转运体1(GLT-1)和GS mRNA表达;Western Blot法检测小鼠大脑皮层和海马GLT-1和GS蛋白表达。结果治疗性给药30 d后,与PTZ组相比,人参皂苷Rb_(1)低、中、高剂量组和VPA组小鼠癫痫发作级别降低,发作潜伏期延长,大脑皮层和海马中Glu含量降低,GS活性升高,GLT-1、GS mRNA表达和蛋白表达增加,海马CA1区FJB阳性细胞数量减少(P<0.05,P<0.01)。结论人参皂苷Rb_(1)治疗性给药能有效缓解癫痫小鼠的发作严重程度,改善神经细胞损伤,发挥抗癫痫作用,其机制可能与上调谷氨酸-谷氨酰胺(Glu-Gln)循环上的关键环节GLT-1和GS的表达,调节Glu代谢有关。Objective To explore the antiepileptic effect of ginsenoside Rb_(1) therapeutic administration and its the mechanism of action.Methods The mouse epilepsy model was established by pentylenetetrazole(PTZ)kindling.The mice were randomly divided into PTZ group,sodium vedproate(VPA)group and low(10 mg·kg^(-1)),middle(20 mg·kg^(-1))and high dose(40 mg·kg^(-1))ginsenoside Rb_(1) groups.Each group was intraperitoneally injected with corresponding medicine for 30 days,once a day.The severity of seizures and the recovery of therapeutic administration in mice were evaluated by observing the seizure stage and seizure latency at different time points after administration.The glutamate(Glu)content and glutamine synthetase(GS)activity in cortex and hippocampus was detected by biochemical method;the degeneration of neurons in hippocampal CA1 was observed by Fluoro-Jade B(FJB)staining;the mRNA expression of glutemate transporter 1(GLT-1)and GS in cortex and hippocampus was detected by reverse transcription-polymerase chain reaction(RT-PCR);the protein expression of GLT-1 and GS in cortex and hippocampus was detected by Western Blot.Results After 30 days therapeutic administration,compared with the PTZ group,the seizure stage decreased,the seizure latency increased,Glu content decreased and GS activity,GLT-1 and GS mRNA and protein expression increased in cortex and hippocampus in VPA group and low,middle and high dose ginsenoside Rb_(1) groups,and the number of FJB positive cells in hippocampal CA1 decreased in VPA group and low,middle and high dose ginsenoside Rb_(1) groups(P<0.05,P<0.01).Conclusion Therapeutic administration of ginsenoside Rb_(1) can alleviate the seizure severity of epileptic mice,improve neuron damage,and play an antiepileptic role.Its mechanism may be related to up-regulating the expression of GLT-1 and GS,which are key links in the glutamate-glutamine(Glu-Gln)cycle,and regulating Glu metabolism.

关 键 词：人参皂苷Rb_(1) 癫痫 谷氨酸(Glu)代谢 谷氨酸-谷氨酰胺(Glu-Gln)循环 

分 类 号：R965[医药卫生—药理学]