Neuroprotective effects of Shaoyao Gancao decoction against excitatory damage in PC12 cells based on the Src-NR2-nNOS pathway  

作　　者：Xiaoxu Fan Hongyan Ma Tiantian Zhou Min Fu Zhiyuan Qiao Yingtong Feng Zhen Wang Yiwei Shen Jingxia Wang 

机构地区：[1]School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China [2]School of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830017,China [3]The Centre for Translational Biology,Research Institute of McGill University Health Centre,Montréal H4A 3J1,Canada

出　　处：《Journal of Traditional Chinese Medical Sciences》2024年第3期293-302,共10页中医科学杂志（英文）

基　　金：supported by the National Natural Science Foundation of China(82074036).

摘　　要：Objective:To explore the neuroprotective effects of the Shaoyao Gancao decoction(SGD)against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulatingγ-aminobutyric acid(GABA)-glutamate(Glu)homeostasis.Methods: N-Methyl-d-aspartic acid(NMDA)was used to establish a PC12 cell excitability injury model.To investigate the neuroprotective effect of SGD,a cell counting kit-8(CCK-8)assay was used to determine PC12 cell viability,Annexin V/Propidium Iodide(Annexin V/PI)double staining was used to determine PC12 cell apoptosis,and Ca^(2+)concentration was observed using laser confocal microscopy.GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions betweenγ-aminobutyric acid(GABA)and NMDA receptors.Additionally,molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway.We analyzed the correlations between the regulatory sites of GABA and NMDA interactions,excitatory neurotoxicity,and brain damage at the molecular level.Results: NMDA excitotoxic injury manifested as a significant decrease in cell activity,increased apoptosis and caspase-3 protein expression,and a significant increase in intracellular Ca^(2+)concentration.Administration of SGD,a GABAA receptor agonist(muscimol),or a GABAB receptor agonist(baclofen)decreased intracellular Ca^(2+)concentrations,attenuated apoptosis,and reversed NMDA-induced upregulation of caspase-3,Src,NMDAR2A,NMDAR2B,and nNOS.Unexpectedly,a GABA_(A)receptor antagonist(bicuculline)and a GABA_(B)receptor antagonist(saclofen)failed to significantly increase excitatory neurotoxicity.Conclusions: Taken together,these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases,but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.

关 键 词：Shaoyao Gancao decoction PC12 N-Methyl-D-aspartic acid(NMDA) γ-aminobutyric acid(GABA) SRC NNOS 

分 类 号：R285[医药卫生—中药学]