摘要
探讨导痰洗心汤对APP/PS1小鼠的治疗效果及其作用机制。将12只APP/PS1雄性小鼠随机分为4组:APP/PS1组、导痰洗心汤低剂量组、导痰洗心汤中剂量组、导痰洗心汤高剂量组,每组3只;C57BL/6野生型小鼠3只作为对照组。采用Morris水迷宫实验评估小鼠学习和记忆能力;通过苏木素-伊红(HE)染色和尼氏染色观察海马组织神经细胞的病理形态变化;利用免疫组化技术检测海马组织中β-淀粉样蛋白(Aβ)_(1-42)表达水平。选取治疗效果显著的导痰洗心汤高剂量组与APP/PS1组进行高通量测序,对测序结果进行差异表达基因、基因本体论(GO)功能分析、京都基因与基因组百科全书(KEGG)通路富集分析和GSVA通路分析,并对部分差异表达基因进行RT-qPCR和Western blot检测,以验证mRNA和蛋白表达情况。与APP/PS1组相比,导痰洗心汤低、中、高剂量可改善APP/PS1小鼠的学习记忆能力,海马组织中CA1区神经病理损伤状况有所改善,神经元数量增加,脑内Aβ_(1-42)沉积减少。差异基因筛选结果中,共有1240个差异基因符合筛选标准,其中上调基因634个,下调基因606个。GO功能富集分析表明,生物过程主要涉及RNA剪接和蛋白质折叠,细胞组分主要为剪接体复合体和核斑点,分子功能主要为未折叠蛋白结合和热休克蛋白结合等。KEGG通路富集分析涉及神经退行性疾病通路、肌萎缩侧索硬化症和剪接复合体等。进一步的GSVA通路富集分析显示,下调通路涉及核因子-κB(NF-κB)介导的肿瘤坏死因子-α(TNF-α)信号通路、紫外响应和未折叠蛋白反应等,上调通路涉及Wnt/β-连环蛋白信号通路等;RT-qPCR和Western blot检测结果显示,与APP/PS1组相比,导痰洗心汤低、中、高剂量组小鼠海马组织中信号转导与转录激活因子3(STAT3)、NF-κB、白细胞介素-6(IL-6)mRNA及蛋白表达降低。以上说明导痰洗心汤可改善APP/PS1小鼠的学习记忆能力,其机制可能与STAT3/NF-κB/IL-6信号通路的调节有关。
This study aims to investigate the therapeutic effect and mechanism of Daotan Xixin Decoction on APP/PS1 mice.Twelve APP/PS1 male mice were randomized into four groups:APP/PS1 and low-,medium-,and high-dose Daotan Xixin Decoction.Three C57BL/6 wild-type mice were used as the control group.The learning and memory abilities of mice in each group were examined by the Morris water maze test.The pathological changes of hippocampal nerve cells were observed by hematoxylin-eosin staining and Nissl staining.Immunohistochemistry was employed to detect the expression ofβ-amyloid(Aβ)_(1-42) in the hippocampal tissue.The high-dose Daotan Xixin Decoction group with significant therapeutic effects and the model group were selected for high-throughput sequencing.The differentially expressed gene(DEG)analysis,Gene Ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,and Gene Set Variation Analysis(GSVA)were performed on the sequencing results.RT-qPCR and Western blot were conducted to determine the mRNA and protein levels,respectively,of some DEGs.Compared with the APP/PS1 group,Daotan Xixin Decoction at different doses significantly improved the learning and memory abilities of APP/PS1 mice,ameliorated the neuropathological damage in the CA1 region of the hippocampus,increased the number of neurons,and decreased the deposition of Aβ_(1-42) in the brain.A total of 1240 DEGs were screened out,including 634 genes with up-regulated expression and 606 genes with down-regulated expression.The GO analysis predicted the biological processes including RNA splicing and protein folding,the cellular components including spliceosome complexes and nuclear spots,and the molecular functions including unfolded protein binding and heat shock protein binding.The KEGG pathway enrichment analysis revealed the involvement of neurodegenerative disease pathways,amyotrophic lateral sclerosis,and splicing complexes.Further GSVA pathway enrichment analysis showed that the downregulated pathways involved nuclear factor-κB(NF-κB)-mediated tumor necrosis factor-α(TNF-α)signaling pathway,UV response,and unfolded protein response,while the up-regulated pathways involved the Wnt/β-catenin signaling pathway.The results of RT-qPCR and Western blot showed that compared with the APP/PS1 group,Daotan Xixin Decoction at different doses down-regulated the mRNA and protein levels of signal transducer and activator of transcription 3(STAT3),NF-κB,and interleukin-6(IL-6)in the hippocampus.In conclusion,Daotan Xixin Decoction can improve the learning and memory abilities of APP/PS1 mice by regulating the STAT3/NF-κB/IL-6 signaling pathway.
作者
陈博伦
卢建政
周欣梅
文晓东
蒋媛静
罗宁
CHEN Bo-lun;LU Jian-zheng;ZHOU Xin-mei;WEN Xiao-dong;JIANG Yuan-jing;LUO Ning(Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,China;Area 1 of the Neurology Department,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,China)
出处
《中国中药杂志》
北大核心
2025年第2期301-312,共12页
China Journal of Chinese Materia Medica
基金
广西自然科学基金项目(2021GXNSFAA196031)。
