摘要
目的探讨艾司氯胺酮(Eske)是否可通过调节高迁移率族蛋白B1(HMGB1)/Toll样受体4(TLR4)信号通路减轻癫痫大鼠神经损伤。方法将大鼠随机分为NC组、Model组、Eske-L组、Eske-H组、Eske-H+rHMGB1(HMGB1激活剂)组。观察大鼠癫痫发作情况;Morris水迷宫实验评估大鼠空间学习记忆能力;Nissl染色观察海马组织形态;TUNEL染色检测神经元凋亡;Fluoro-Jade B染色检测损伤神经元;免疫荧光染色检测神经元特异性核蛋白(NeuN)标记的神经元核抗原;ELISA检测TNF-α、IL-10、IL-1β、γ-氨基丁酸(GABA)水平;蛋白印迹法检测Cleaved-caspase-3、HMGB1、TLR4、NF-κB和p-NF-κB蛋白表达水平。结果与NC组相比,Model组大鼠海马CA3区组织严重损伤,癫痫发作时间和潜伏期延长,Racine评分、TNF-α和IL-1β水平、神经元凋亡率、损伤神经元数量、Cleavedcaspase-3、HMGB1、TLR4和p-NF-κB/NF-κB蛋白表达水平升高,大鼠滞留时间缩短,穿越平台次数减少,IL-10和GABA水平、NeuN神经元数量降低(P<0.05);与Model组比较,Eske-L组和Eske-H组大鼠海马CA3区组织损伤程度减轻,癫痫发作时间和潜伏期缩短,Racine评分、TNF-α和IL-1β水平、神经元凋亡率、损伤神经元数量、Cleaved-caspase-3、HMGB1、TLR4和p-NF-κB/NF-κB蛋白表达水平降低,滞留时间延长,穿越平台次数增加,IL-10和GABA水平、NeuN神经元数量升高(P<0.05);rHMGB1可减弱Eske对癫痫大鼠神经损伤的改善作用(P<0.05)。结论Eske可能通过抑制HMGB1/TLR4通路来降低癫痫大鼠炎性反应和神经元凋亡,减轻神经损伤,改善其认知功能。
Objective To explore whether esketamine(Eske)can alleviate nerve injury in epileptic rats by regulating the High mobility group box protein B1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway.Methods The rats were randomly allocated into five groups:NC group,Model group,Eske low-dose group(Eske-L),Eske high-dose group(Eske-H),and Eske-H plus rHMGB1(HMGB1 activator)group.Observe the epileptic seizures in rats;Evaluate the spatial learning and memory abilities of rats by Morris water maze experiment;Observation of hippocampal tissue morphology using Nissl staining;Detect the apoptosis of hippocampal neurons by TUNEL;Fluoro-Jade B staining was used to detect damaged neurons;Detect neuronal nuclear antigens labeled with neuron-specific nuclear protein(NeuN)by Immunofluorescence staining;The levels of TNF-α,IL-10,IL-1β,and GABA were detected by ELISA;the expression levels of Cleaved-caspase-3,HMGB1,TLR4,NF-κB,and p-NF-κB proteins were detected by protein immunoblotting.Results For the NC group,the tissue in the hippocampal CA3 area of rats in the Model group was severely damaged,and the duration and latency of epileptic seizures were prolonged.The Racine score,TNF-α,IL-1βlevels,neuronal apoptosis rate,the number of injured neurons,the protein expression levels of Cleaved-caspase-3,HMGB1,TLR4 and p-NF-κB/NF-κB increased,the retention time shortened,and the number of crossing platforms decreased.The levels of IL-10 and GABA and the number of NeuN neurons decreased(P<0.05).For the Model group,the degree of tissue injury in the hippocampal CA3 area of rats in the Eske-L group and the Eske-H group was reduced,and the duration and latency of epileptic seizures were shortened.The Racine score,TNF-α,IL-1βlevels,neuronal apoptosis rate,the number of injured neurons,the protein expression levels of Cleaved-caspase-3,HMGB1,TLR4 and p-NF-κB/NF-κB decreased,the duration of stay has been extended,and the number of crossings across the platform has increased.The levels of IL-10 and GABA and the number of NeuN neurons increased(P<0.05).rHMGB1 can weaken the improvement effect of Eske on nerve injury in epileptic rats(P<0.05).Conclusion Eske may reduce the inflammatory response and neuronal apoptosis of epileptic rats,alleviate neural injury and improve their cognitive function by inhibiting the HMGB1/TLR4 pathway.
作者
赵华英
张冰
刘步云
艾田妹
国义民
Zhao Huaying;Zhang Bing;Liu Buyun;Ai Tianmei;Guo Yimin(Department of Neurology,Hengshui Second Peoples Hospital,Hebei O53000,China)
出处
《脑与神经疾病杂志》
2026年第1期57-63,共7页
Journal of Brain and Nervous Diseases
基金
2017年衡水市科技计划(医药卫生部分)项目(2017014013C-22)。
