草果知母汤对戊四唑诱导癫痫大鼠海马神经元的保护作用及RECK/GFAP/AKT信号通路的调节作用  

Protective Effect of Caoguo Zhimu Decoction on Hippocampal Neurons and Regulation of RECK/GFAP/AKT Signal Pathway in Epileptic Rats Induced by Pentylenetetrazol

作　　者：张高炼[1] 郭建辉 曾敬 李敏[1] 陈锐[1] 张晓宁[1] 刘姗姗[1] 梁韡斌 ZHANG Gaolian;GUO Jianhui;ZENG Jing;LI Min;CHEN Rui;ZHANG Xiaoning;LIU Shanshan;LIANG Weibin(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine, Nanning 530023,Guangxi,China)

机构地区：[1]广西中医药大学第一附属医院,南宁530023

出　　处：《中西医结合心脑血管病杂志》2022年第5期857-862,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金项目(No.81660782)。

摘　　要：目的研究草果知母汤对戊四唑诱导癫痫大鼠海马神经元的保护作用及反转录富含半胱氨酸蛋白(RECK)/胶质纤维酸性蛋白(GFAP)/蛋白激酶B(AKT)信号通路的调节作用。方法将90只雄性SD大鼠随机分为正常对照组、癫痫模型组、草果知母汤低剂量组、草果知母汤中剂量组、草果知母汤高剂量组及阳性对照组,每组15只,大鼠采用腹腔注射戊四唑方法建立癫痫大鼠模型,造模成功后,草果知母汤低、中、高剂量组分别给予2.5 mL/kg、5.0 mL/kg、10.0 mL/kg的草果知母汤灌胃,阳性对照组给予丙戊酸钠,每日1次,连续给药4周,实时定量聚合酶链式反应(RT-qPCR)测定海马组织Bax、Bcl-2、Caspase-3 mRNA水平,尼氏染色法观察大鼠海马神经元尼氏小体水平,脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)染色法检测大鼠海马神经元凋亡水平,免疫蛋白印迹法(Western Blot)检测大鼠海马组织RECK、GFAP、磷酸化蛋白激酶B(p-AKT)、AKT水平。结果与癫痫模型组比较,草果知母汤各剂量组大鼠海马组织Bcl-2 mRNA水平、RECK及p-AKT水平升高,Bax、Caspase-3 mRNA水平、GFAP蛋白水平及海马神经元凋亡数降低,差异均有统计学意义(P<0.05),且各项指标变化表现出剂量依赖性,海马神经元尼氏小体染色明显改善。结论草果知母汤能够降低戊四唑诱导的癫痫大鼠海马神经元凋亡,调节凋亡相关基因表达水平,其机制可能与调节RECK/GFAP/AKT信号通路有关。Objective To study the protective effect of Caoguo Zhimu Decoction(CGZM)on hippocampal neurons and regulation of reversion-inducing cysteine-rich protein with Kazal motif(RECK)/glial fibrillary acidic portein(GFAP)/protein kinase B(AKT)signal pathway in pentylenetetrazol-induced epileptic rats.Methods Ninety rats were randomly divided into normal control group,epileptic model group,low dose CGZM group,middle dose CGZM group,and high dose CGZM group,and positive control group,with 15 rats in each group.The epileptic rat model was established by intraperitoneal injection of pentylenetetrazol.After successful establishment,the rats in low dose CGZM group,middle dose CGZM group,and high dose CGZM group were given CGZM of 2.5 mL/kg,5.0 mL/kg,and 10.0 mL/kg,respectively,and the rats in positive control group were given sodium valproate once a day for 4 weeks.The levels of Bax,Bcl-2,and Caspase-3 mRNA in hippocampus were measured by real-time quantitative PCR.Nissl bodies of hippocampal neurons were detected by Nissl staining.Apoptosis of hippocampal neurons was detected by terminal deoxynucl neotidyl transferase mediated dUTP nick end labelling(TUNEL)staining.Western blot was used to detect the levels of RECK,GFAP,phosphorylated AKT(p-AKT),and AKT in hippocampal tissue.Results Compared with epilepsy model group,the levels of Bcl-2 mRNA,RECK,and p-AKT in hippocampal tissue were increased,the levels of Bax,Caspase-3 mRNA,and GFAP protein and the number of apoptotic hippocampal neurons decreased,and the changes of various indexes were dose-dependent.The Nissl body staining of hippocampal neurons was improved significantly in each dose group.Conclusion CGZM can reduce the apoptosis of hippocampal neurons and regulate the expression of apoptosis-related genes in epileptic rats induced by pentylenetetrazol.The mechanism may be related to the regulation of RECK/GFAP/AKT signal pathway.

关 键 词：癫痫 草果知母汤 神经元保护 反转录富含半胱氨酸蛋白 胶质纤维酸性蛋白 蛋白激酶B 实验研究 

分 类 号：R73[医药卫生—肿瘤]