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向静,江姗姗,唐红,汪红娟,李展富,王瑶,谢灿明,田浩梅,陈楚淘.针刺调控脑缺血再灌注损伤大鼠海马区环状RNA_009775表达的研究[J].湖南中医药大学学报英文版,2023,43(10):1838-1845.[Click to copy
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| 针刺调控脑缺血再灌注损伤大鼠海马区环状RNA_009775表达的研究 |
| 向静,江姗姗,唐红,汪红娟,李展富,王瑶,谢灿明,田浩梅,陈楚淘 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 研究针刺对脑缺血再灌注损伤（cerebral ischemia reperfusion injury, CIRI）大鼠海马区环状RNA_009775表达的影响，探索针刺对CIRI可能的作用机制。方法 将39只SD大鼠随机分为假手术组、模型组、针刺组，每组13只。除假手术组外，其余两组均采用线栓法制备CIRI模型。造模后，针刺组大鼠捆绑固定行针刺干预，间隔12 h进行1次，共7次；假手术组与模型组大鼠仅捆绑不针刺。Garcia评分法、TTC染色法评估针刺疗效；基因芯片技术筛选针刺组/模型组、模型组/假手术组核心共同差异表达circRNA；qRT-PCR法对芯片筛选结果进行验证；TargetScan软件预测核心circRNA的下游miRNA及mRNA，构建核心circRNA的调控网络，并对mRNA进行基因本体功能富集分析；Western blot法验证神经元核抗原抗体（neuron specific nuclear protein, NEUN）表达量。结果 干预后，与假手术组相比，模型组大鼠Garcia评分、NEUN和circRNA_009775表达量显著降低（P＜0.01），脑梗死面积比显著升高（P＜0.01）；与模型组相比，针刺组脑梗死面积比显著降低（P＜0.01），Garcia评分、NEUN和circRNA_009775表达量显著升高（P＜0.05或P＜0.01）。芯片筛选核心共同差异表达circRNA，经qRT-PCR验证circRNA_009775表达趋势与芯片结果一致，结合靶关系预测，构建了含有1个circRNA、12个miRNA、31个mRNA的调控网络。基因本体功能富集分析显示mRNA主要涉及神经系统发育、神经元生成等生物过程。结论 针刺具有改善CIRI大鼠神经功能缺损、降低脑梗死面积、部分减轻海马区神经元损伤的作用，其机制可能与调控circRNA_009775介导的潜在调控网络、促进神经系统发育、神经元生成等生物过程有关。 |
| 关键词: 脑缺血再灌注损伤 针刺 海马区 环状RNA 基因芯片 神经元 督脉 |
| DOI：10.3969/j.issn.1674-070X.2023.10.013 |
| Received:June 13, 2023 |
| 基金项目:国家自然科学基金项目（82274662）；湖南省自然科学基金项目（2021JJ30490）；长沙市科技局自然科学基金项目（kq2014094）；湖南省研究生科研创新项目（2022CX103）。 |
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| Acupuncture regulating the expression of circRNA_009775 in hippocampus of rats with cerebral ischemia- reperfusion injury |
| XIANG Jing,JIANG Shanshan,TANG Hong,WANG Hongjuan,LI Zhanfu,WANG Yao,XIE Canming,TIAN Haomei,CHEN Chutao |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To study the effects of acupuncture on the expression of circRNA_009775 in hippocampus of rats with cerebral ischemia-reperfusion injury (CIRI), and to explore the possible mechanism of action of acupuncture on CIRI. Methods A total of 39 SD rats were randomly assigned into sham-operated, model, and acupuncture groups, with 13 rats in each group. Except for the sham-operated group, CIRI model was established using the modified Zea Longa method in the other two groups. After modeling, the rats in acupuncture group were bundled for acupuncture intervention, once every 12 hours, 7 times in total; the rats in sham-operated group and model group were only bundled without acupuncture. Garcia scoring and TTC staining were used to evaluate the efficacy of acupuncture; gene chip microarray analysis was used to screen the core common differentially expressed circRNAs (co-DEcircRNAs) in acupuncture/model and model/sham-operated groups; qRT-PCR was used to validate the microarray results; TargetScan software was applied to predicting the downstream miRNAs and mRNAs of the core co-DEcircRNAs and constructing ceRNA network, and gene ontology function enrichment analysis was performed for the target mRNAs; Western blot was performed to verify the expression level of neuron specific nuclear protein (NeuN). Results After the intervention, compared with sham-operated group, the Garcia score and expression levels of NeuN and circRNA_009775 of rats in model group significantly decreased (P＜0.01), and the ratio of cerebral infarction area significantly increased (P＜0.01). Compared with model group, the ratio of cerebral infarction area of rats in acupuncture group was significantly reduced (P＜0.01), and the Garcia score and expression levels of NeuN and circRNA_009775 were significantly elevated (P＜0.05 or P＜0.01). The core co-DEcircRNAs were screened out by gene chip microarray analysis, and the expression trend of circRNA_009775 verified by qRT-PCR was consistent with the gene microarray results. Based on the target relation prediction, a ceRNA network containing 1 circRNA, 12 miRNAs, and 31 mRNAs was constructed. Gene ontology functional enrichment analysis showed that the target mRNAs were mainly involved in biological processes such as neurological development and neuron generation. Conclusion Acupuncture has the effects of improving neurological deficits, reducing the area of cerebral infarction, and partially alleviating neuronal damage in hippocampus of CIRI rats, the mechanism of which may be related to regulating circRNA_009775-mediated potential ceRNA network to promote biological processes such as neurological development and neuron generation. |
| Key words: cerebral ischemia-reperfusion injury acupuncture hippocampus circRNA gene chips neuron the Du meridian |
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