针刺研究

2021, v.46(09) 782-788+808

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基于转录组测序技术探讨蓝斑核参与针刺抗心肌缺血的分子机制
Molecular mechanism of locus coeruleus involved in acupuncture anti-myocardial ischemia based on transcriptome sequencing

吴欣;王晨凯;左海燕;瞿巧钰;童艳;王堃;吴生兵;周美启;
WU Xin;WANG Chen-kai;ZUO Hai-yan;QU Qiao-yu;TONG Yan;WANG Kun;WU Sheng-bing;ZHOU Mei-qi;Graduate School,Anhui University of Chinese Medicine;School of Acupuncture-moxibustion and Massage,Anhui University of Chinese Medicine;Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine;Research Institute of Acupuncture-moxibustion and Meridian,Anhui University of Chinese Medicine;Bozhou Institute of Chinese Medicine,Anhui Academy of Traditional Chinese Medicine;

摘要(Abstract):

目的:探讨蓝斑核参与针刺抗心肌缺血的分子机制。方法:将雄性SD大鼠随机分成伪手术组、模型组、电针组、电针+损毁组,每组6只。采用冠状动脉左前降支结扎法复制急性心肌缺血模型。电针组在"神门"-"通里"段给予电针,强度1 mA,频率2 Hz/15 Hz,每次30 min,连续3 d;电针+损毁组在病毒损毁双侧蓝斑核的基础上复制急性心肌缺血模型,再给予和电针组同样的电针治疗。用酶联免疫法检测血清天门冬氨酸氨基转移酶(AST),采用转录组测序技术检测各组大鼠心脏的基因表达谱,筛选差异表达基因,并进行基因本体论(GO)功能分类以及京都基因与基因组百科全书(KEGG)代谢通路富集分析。结果:与伪手术组比较,模型组大鼠血清AST显著升高(P<0.01);与模型组比较,电针组血清AST显著降低(P<0.01);与电针组比较,电针+损毁组血清AST显著升高(P<0.05)。与伪手术组比较,模型组共筛选出1 138条差异表达基因。与模型组比较,电针组共筛选出1 330条差异表达基因。与电针组比较,电针+损毁组共筛选出804条差异表达基因。其中蓝斑核参与针刺抗心肌缺血所调节的差异基因共218条,GO功能分类分析显示在生物过程中这些差异基因主要涉及细胞过程、代谢过程和生物调节功能。KEGG代谢通路分析显示这些差异基因富集于硫中继系统、硫胺素代谢、谷胱甘肽代谢、C5分支二元酸新陈代谢、细胞黏附分子和Th1和Th2细胞分化等。结论:蓝斑核参与针刺抗心肌缺血的分子机制可能与硫中继系统、硫胺素代谢、谷胱甘肽代谢、C5分支二元酸新陈代谢、细胞黏附分子和Th1和Th2细胞分化以及相关基因密切相关。
Objective To explore the molecular mechanism of locus coeruleus(LC) involved in electroacupuncture(EA) anti myocardial ischemia. Methods Twenty-four SD rats were randomly divided into sham-operation, model, EA and EA +lesion groups, with 6 rats in each group. The acute myocardial ischemia(AMI) model was established by ligation of the left anterior descending branch of coronary artery. EA(2 Hz/15 Hz, 1 mA) was applied to bilateral "Shenmen"(HT7)-"Tongli"(HT5) and the middle-point between HT7 and HT5 for 30 min, once daily for 3 days. For rats of the EA +lesion group, the virus(300 nL) was injected into bilateral LC before EA treatment. Serum aspartate aminotransferase(AST) was detected by ELISA. The gene expression profiles of rat heart were detected by transcriptome sequencing, the differentially expressed genes were screened, and Gene Ontology(GO) functional classification and Kyoto Encyclopedia of genes and genomes(KEGG) metabolic pathway enrichment analysis were performed. Results Compared with the sham-operation group, serum AST content was significantly increased in the model group(P<0.01). Following the intervention, serum AST was significantly reduced in the EA group(P<0.01), while the serum AST in the EA + lesion group was significantly higher compared with the EA group(P<0.05). Differential expression analysis showed that 1 138 differentially expressed genes were screened out between the model group and the sham-operation group, 1 330 differentially expressed genes between model and EA group, and 804 differentially expressed genes between EA and EA + lesion group. Among them, 218 differential genes were involved in the regulation of EA anti-myocardial ischemia in LC. GO functional classification analysis showed that these differentially expressed genes mainly involved in cell processes, metabolic processes and biological regulation in biological processes. KEGG pathway analysis showed that these differentially expressed genes were enriched in sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation. Conclusion EA intervention has a positive effect in anti-myocardial ischemia, which may be related to the sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation involved in LC.

关键词(KeyWords): 转录组;急性心肌缺血;心肌组织;电针
Transcriptome;Acute myocardial ischemia;myocardial tissue;Electroacupuncture

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81674058、81273858、81804191、82004462)

作者(Author): 吴欣;王晨凯;左海燕;瞿巧钰;童艳;王堃;吴生兵;周美启;
WU Xin;WANG Chen-kai;ZUO Hai-yan;QU Qiao-yu;TONG Yan;WANG Kun;WU Sheng-bing;ZHOU Mei-qi;Graduate School,Anhui University of Chinese Medicine;School of Acupuncture-moxibustion and Massage,Anhui University of Chinese Medicine;Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine;Research Institute of Acupuncture-moxibustion and Meridian,Anhui University of Chinese Medicine;Bozhou Institute of Chinese Medicine,Anhui Academy of Traditional Chinese Medicine;

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DOI: 10.13702/j.1000-0607.200722

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