基于NMDAR1-PSD-nNOS通路探讨栝楼桂枝颗粒对神经元兴奋性毒性损伤后突触可塑性的影响  

Effect of Gualou Guizhi Granule on Synaptic Plasticity after Neuronal Excitotoxic Injury Based on NMDAR1-PSD-nNOS Pathway

作　　者：冯怡 李雪珍 李亚男 陈雯婷 罗书萍 张玉琴[1] FENG Yi;LI Xuezhen;LI Yanan;CHEN Wenting;LUO Shuping;ZHANG Yuqin(Pharmacy College of Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《福建中医药》2025年第1期17-21,29,共6页Fujian Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81803768);福建省自然科学基金面上项目(2024J01127);福建中医药大学基础类学科科研提升计划项目(XJC2022009)。

摘　　要：目的基于NMDAR1-PSD-nNOS通路探讨栝楼桂枝颗粒(GLGZG)对神经元兴奋性毒性损伤后突触可塑性的影响,为GLGZG治疗缺血性脑卒中提供实验依据。方法取新生大鼠的原代海马神经元,分为对照组、模型组、低剂量组、中剂量组,建立N-甲基-D-天冬氨酸(NMDA)诱导神经元兴奋性毒性损伤模型,对照组加入不含有NMDA的完全培养基进行培养,模型组用含有200μmol/L NMDA的完全培养基诱导损伤6 h,低、中剂量组分别予以含有100、200μg/mL GLGZG溶液的完全培养基干预24 h后,再更换为含有200μmol/L NMDA的完全培养基诱导损伤6 h。干预后采用电子显微镜观察4组神经元形态,MTT法检测4组神经元存活率,乳酸脱氢酶(LDH)法检测4组神经元LDH水平,qPCR检测4组神经元NeuN、GAP-43、PSD-95、NMDAR1 mRNA转录水平,Western blot检测4组神经元NeuN、GAP-43、PSD-95、MAP-2、Synapsin-1、NMDAR1、nNOS蛋白相对表达量。结果与对照组比较,模型组神经元形态异常,神经元存活率降低,LDH水平升高,NeuN、GAP-43、PSD-95、NMDAR1 mRNA转录水平均显著降低(P<0.05),NeuN、GAP-43、PSD-95、MAP-2、Synap⁃sin-1、NMDAR1、nNOS蛋白相对表达量均降低(P<0.05)。与模型组比较,低、中剂量组神经元形态明显改善且神经元存活率显著提高,LDH水平降低(P<0.05);低剂量组NeuN、NMDAR1 mRNA转录水平以及NeuN、GAP-43、PSD-95、Synapsin-1蛋白相对表达量均上升(P<0.05),中剂量组NeuN、GAP-43、PSD-95、NMDAR1 mRNA转录水平以及NeuN、GAP-43、PSD-95、Synapsin-1、MAP-2、NMDAR1、nNOS蛋白相对表达量均提高(P<0.05)。与低剂量组比较,中剂量组LDH水平以及MAP-2、Synapsin-1蛋白相对表达量均升高(P<0.05)。结论GLGZG能够减轻NMDA诱导的神经元兴奋性毒性损伤,可能与GLGZG可通过调控NMDAR1-PSD-nNOS通路、改善神经元突触可塑性有关。Objective:To study the effect of Gualou Guizhi Granule(GLGZG)on synaptic plasticity after neuronal excitotoxicity injury based on the NMDAR1-PSD-nNOS pathway,and to provide an experimental basis for the treatment of ischemic stroke with GLGZG.Methods:Primary hippocampal neurons were isolated from newborn rats and divided into control group,model group,low-dose group,and medium-dose group.An N-methyl-D-aspartate(NMDA)-induced neuronal excitotoxic injury model was established.The control group was cultured in complete medium without NMDA.The model group was induced with complete medium containing 200μmol/L NMDA for 6 hours.The low and medium-dose groups were intervened with complete medium containing 100 and 200μg/mL GLGZG solution,respectively,for 24 hours before being exposed to 200μmol/L NMDA for 6 hours.After intervention,neuronal morphology was observed using an electron microscope.Neuronal survival rate was detected by the MTT assay,lactate dehydrogenase(LDH)levels were measured using the LDH method,and the transcription levels of NeuN,GAP-43,PSD-95,and NMDAR1 mRNA in neurons were detected by qPCR.The relative expression levels of NeuN,GAP-43,PSD-95,MAP-2,Synapsin-1,NMDAR1,and nNOS proteins were detected by Western blot.Results:Compared with the control group,neuronal morphology of the model group was abnormal,neuronal survival rate and transcription levels of NeuN,GAP-43,PSD-95,and NMDAR1 mRNA decreased,as well as relative expression levels of NeuN,GAP-43,PSD-95,MAP-2,Synapsin-1,NMDAR1,and nNOS proteins(P<0.05),but LDH level increased(P<0.05).Compared with the model group,the low and medium-dose groups showed significant improvement in neuronal morphology and neuronal survival rate increased,with LDH levels reducing(P<0.05).The transcription levels of NeuN and NMDAR1 mRNA,as well as the relative expression levels of NeuN,GAP-43,PSD-95,and Synapsin-1 proteins,were increased in the low-dose group(P<0.05).In the medium-dose group,the transcription levels of NeuN,GAP-43,PSD-95,and NMDAR1 mRNA,as well as the r

关 键 词：神经元损伤 脑卒中 神经可塑性 栝楼桂枝颗粒 

分 类 号：R74[医药卫生—神经病学与精神病学]