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基于转录组测序技术研究川芎嗪对急性脊髓损伤模型大鼠基因表达的影响
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| 篇名： | 基于转录组测序技术研究川芎嗪对急性脊髓损伤模型大鼠基因表达的影响 |
| TITLE： | Study on the Effects of Ligustrazine on Gene Expression of Acute Spinal Cord Injury Model Rats Based on Transcriptome Sequencing |
| 摘要： | 目的：探讨川芎嗪对急性脊髓损伤（SCI）模型大鼠基因表达的影响。方法：将雄性SD大鼠随机分为假手术组（A组，6只），模型组（B组，不同时间点各6只，共12只）和川芎嗪干预组（C组，不同时间点各6只，共12只）。B、C组大鼠参照改良Allen’s法复制急性SCI模型。造模后，C组大鼠腹腔注射川芎嗪100mg/kg，A、B组大鼠腹腔注射等容生理盐水，每日1次，持续7d或14d（分别记为B7d、B14d组和C7d、C14d组）。分别于造模前和造模后7、14d对各组大鼠进行BBB评分，并行脊髓标本苏木精-伊红、尼氏染色观察；采用转录组测序技术分析A组与B组、B组与C组大鼠脊髓标本中的差异表达基因（DEGs），借助基因本体（GO）数据库和KEGG数据库进行GO和KEGG信号通路富集分析。结果：与A组比较，B组大鼠造模后7、14d的BBB评分均显著降低（P＜0.05），其脊髓组织中的神经细胞和尼氏体数量明显减少；与B组比较，C组大鼠上述时间点的BBB评分均显著升高（P＜0.05），其脊髓组织中的神经细胞和尼氏体数量有所增多。A组和B7d组、A组和B14d组、B7d组和C7d组、B14d组和C14d组大鼠的DEGs分别有886、1404、70、66个，组间表达变化趋势相反的基因包括Ncmap、Prx、Gabrq、Gabrg2等。各组间DEGs富集的细胞部位、分子功能、生物学过程各有不同，主要涉及溶泡、溶酶体、质膜部分、同型蛋白结合、免疫应答、离子通道活性、免疫反应（A组和B组），基底外侧质膜、外脱氧核糖核酸酶活性、对干扰素γ的反应（B7d组和C7d组），以及细胞外区、受体调节活性、含酚化合物代谢过程（B14d组和C14d组）等；同时，DEGs富集于细胞因子-细胞因子受体相互作用（A组和B组），细胞黏附分子、补体和凝血级联、Hedgehog信号通路（B7d组和C7d组），逆行内源性大麻素信号、神经活性配体-受体相互作用、过氧化物酶体增殖物激活受体信号通路、γ-氨基丁酸（GABA）能突触（B14d组和C14d组）等信号通路。结论：川芎嗪对急性SCI模型大鼠的保护作用可能与其参与炎症反应、免疫应答/调节、离子通道、细胞因子-细胞因子受体相互作用、神经活性配体-受体相互作用、GABA能突触活性调节等有关。 |
| ABSTRACT： | OBJECTIVE：To investigate the effects of ligustrazine on ge ne expression of acute spinal cord injury （SCI）model rats. METHODS ：Male SD rats were randomly divided into sham operation group （group A ，6 rats），model group （group B ，6 rats at each time point ，12 rats in total ）and ligustrazine intervention group （group C ，6 rats at each time point ，12 rats in total ）. Acute SCI model was established by modified Allen ’s method in group B and C. After modeling ，group C was given ligustrazine 100 mg/kg intraperitoneally ，while group A and B were given constant volume of normal saline intraperitoneally ，once a day ，for consecutive 7 d or 14 d（i.e. group B 7d and B 14d，group C 7d and C 14d）. BBB scoring was conducted in each group before modeling ， 7 and 14 days after modeling. HE and Nissl staining observation were also carried out for spinal cord specimen. The differentially expressed genes （DEGs）between group A and group B ，group B and group C were analyzed by transcriptome sequencing. The enrichment of Gene Ontology （GO）and KEGG signaling pathway was analyzed by GO database and KEGG database. RESULTS ： Compared with group A ，BBB scores of group B were decreased significantly 7 d and 14 d after modeling （P＜0.05），and the number of nerve cells and Nissl body in spinal cord tissue were decreased. Compared with group B ，BBB scores of group C were increased significantly at above time points （P＜0.05），and the number of nerve cells and Nissl body in spinal cord tissue were increased. The numbers of DEGs of group A and group B 7 d， group A and group B 14d，group B 7d and group C 7d，group B 14d FAA380076） and group C 14 d were 886，1 404，70，66，respectively. The genes with opposite expression trend included Ncmap，Prx， Gabrq， Gabrg2， etc. The enrichment cell component ， molecular function ，biological process of DEGs were different 630179114@qq.com in each group ，mainly involving lyocytosis ，lysosome，plasmamembrane，homotype protein binding ，immune response ，ion channel activity ，immune response （group A and B ）；basolateral plasma membrane ，exodeoxyribonuclease activity ，response to INF-γ （group B 7 d and C 7 d）；extracellular domain ，receptor regulatory activity ，phenolic compound metabolism process （group B 14 d and C 14 d）. DEGs enriched in cytokine-cytokine receptor interaction（group A and B ）；CAMs，complement and coagulation cascades and Hedgehog signaling pathway （group B 7d and C 7d）； retrograde endocannabinoid signaling ，neuroactive ligand-receptor interaction ，PPAR signaling pathway ，GABA ergic synapse （group B 14 d and C 14 d），etc. CONCLUSIONS ：Protective effect of ligustrazine on acute SCI model rats may be associated with inflammatory response ，immune response/regulation ，neuron ion channel ，cytokine-cytokine receptor interaction ，neuroactive ligand-receptor interaction and regulation of GABA ergic synapse activity . |
| 期刊： | 2020年第31卷第11期 |
| 作者： | 张毅，祁文，吴迪，谢旻成 |
| AUTHORS： | ZHANG Yi，QI Wen，WU Di，XIE Mincheng |
| 关键字： | 川芎嗪；急性脊髓损伤；转录组测序技术；差异表达基因；基因本体；KEGG信号通路；机制；大鼠 |
| KEYWORDS： | Ligustrazine；Acute spinal cord injury ；Transcriptome sequencing ；Differentially expressed gene ；Gene Ontology ； |
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