抗痫增智颗粒调控TLR4/HMGB1/p-IκB-α信号通路保护癫痫幼鼠海马神经元的分析  

Kangxianzengzhi granules regulate the TLR4/HMGB1/p-IκB-αsignaling pathway to protect hippocampal neurons in juvenile epileptic rats

作　　者：张英英[1] 单海军[1] 娄元俊[1] 郭鑫[1] ZHANG Yingying;SHAN Haijun;LOU Yuanjun;GUO Xin(Children’s Encephalopathy Rehabilitation Department in Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南省中医院儿童脑病康复科,郑州450002

出　　处：《中国比较医学杂志》2020年第10期92-98,共7页Chinese Journal of Comparative Medicine

摘　　要：目的观察抗痫增智颗粒对癫痫幼鼠海马神经元的保护作用,并探讨对调控Toll样受体4(TLR4)/高迁移率族蛋白1(HMGB1)/磷酸化核因子抑制蛋白α(p-IκB-α)信号通路的调控作用。方法建立癫痫模型,阳性组予以托吡酯5.9 mg/kg溶于2 mL/100 g生理盐水中灌胃,低浓度组、中浓度组、高浓度组予以抗痫增智颗粒药液灌胃,抗痫增智颗粒质量分别是1.28 g/kg、2.56 g/kg、5.12 g/kg,模型组和正常组予以生理盐水灌胃,均每天1次,干预4周。Racine分级标准和Morris水迷宫实验评价干预前后行为学,学习/记忆能力变化;苏木素-伊红(HE)染色观察海马区病理变化;逆转录实时定量聚合酶链反应(qRT-PCR)检测海马区TLR4、HMGB1 mRNA表达,蛋白质免疫印迹法(Western blot)检测海马区TLR4、HMGB1、IκB-α蛋白的相对表达量及p-IκB-α。结果干预后阳性组和3浓度组行为学Racine分级、学习/记忆能力指标均较干预前显著改善;干预后模型组海马神经元病理严重变化,阳性组和3浓度组均有所改善;阳性组和3浓度组海马区神经元损伤减轻;模型组TLR4、HMGB1 mRNA、TLR4、HMGB1蛋白的相对表达量、p-IκB-α水平均较正常组显著升高(P<0.01),阳性组和3浓度组均较模型组显著下降(P<0.01);中浓度组和阳性组上述指标差异均无统计学意义(P>0.05),且抗痫增智颗粒的作用呈浓度依赖性。结论抗痫增智颗粒可改善癫痫幼鼠的行为学与学习/记忆能力,减轻海马神经元变性和坏死,且高浓度药物的作用优于托吡酯,推测是通过调控TLR4/HMGB1/p-IκB-α实现的。Objective To observe the protective effect of Kangxianzengzhi granules on hippocampal neurons of juvenile epileptic rats and explore its regulating effect on Toll-like receptor 4(TLR4)/high mobility positive group box 1(HMGB1)/phosphorylated nuclear factor inhibitory proteinα(p-IκB-α)signaling pathways.Methods Epilepsy models were established.In the positive group,5.9 mg/kg topiramate was dissolved in 2 mL/100 g normal saline,and in the low-,medium-,and high-concentration groups,1.28,2.56,and 5.12 g/kg Kangxianzengzhi granules were infused into the stomach,respectively.Model and normal groups had normal saline infused into the stomach once a day for 4 weeks.The changes in behavior and learning/memory abilities before and after the interventions were evaluated by the Racine grading standard and Morris water maze.Hematoxylin-eosin staining was used to observe pathological changes of the hippocampus.Expression of TLR4 and HMGB1 mRNAs in the hippocampus as detected by Real-time quantitative reverse transcription polymerase chain reaction,and relative expression of TLR4,HMGB1,IκB-α,and P-IκB-αwas detected by Western blot.Results After the interventions,the behavioral Racine grade and learning/memory ability indexes of the positive group and three concentration groups were improved significantly compared with those before treatments.After interventions,pathological changes of hippocampal neurons in the model group were serious,which were improved in the positive group and three concentration groups.Damage of hippocampal neurons in the positive group and three concentration groups was alleviated.The relative expression of TLR4 and HMGB1 mRNAs and proteins and the level of P-IκB-αin the model group were significantly higher than those in the normal group(P<0.01),and those in the positive group and three concentration groups were significantly lower than those in the model group(P<0.01).There was no significant difference between the two groups(P>0.05),and the effect of Kangxianzengzhi granules was dependent on c

关 键 词：抗痫增智颗粒 TOLL样受体4 高迁移率族蛋白1 癫痫 海马神经元 

分 类 号：R-33[医药卫生]