CAJ | 학술논문

目的研究电刺激迷走神经对大鼠缺血再灌注心肌基因表达的影响以从基因水平探讨心肌保护机制.方法将20只SD雄性大鼠按随机数字表法分成2组,缺血再灌注组(I/R组)和迷走神经电刺激组(STM组),每组10只.通过结扎冠状动脉左前降支30 min后,松开缝线使冠状动脉再通120 min的方法制备心肌缺血再灌注模型.STM组在缺血再灌注前10 min开始以5V、2 ms、1 Hz强度持续电刺激颈部左侧迷走神经20 Bin.再灌注120 min后将每组按随机数字表法分为2部份,一部份取左心室心肌组织测定心肌梗死面积;一部分取左心室缺血区心肌组织成功提取RNA后,获得心肌基因表达谱.运用GeneSpring软件筛选出两组间差异表达显著的基因并进行分析,对其中具有代表意义的基因进行实时定量逆转录多聚酶链反应(Real-time PCR)验证.结果 (1)STM组心肌梗死区面积较I/R组明显缩小(P＜0.05).(2)基因表达谱筛选得到186个差异表达显著的基因或探针,基因本体分析(GO)显示主要影响了3类基因的改变,上调代表基因与心肌保护有关;下调代表基因与炎症有关.(3)Real-time PCR结果和基因芯片表达的结果相符.结论电刺激迷走神经对缺血再灌注心肌具有保护作用,其心肌基因表达谱发生了很大的变化,可能通过兴奋胆碱能抗炎通路来减轻和抑制心肌缺血再灌注损伤.
목적 연구전자격미주신경대대서결혈재관주심기기인표체적영향이종기인수평탐토심기보호궤제.방법 장20지SD웅성대서안수궤수자표법분성2조,결혈재관주조(I/R조)화미주신경전자격조(STM조),매조10지.통과결찰관상동맥좌전강지30 min후,송개봉선사관상동맥재통120 min적방법제비심기결혈재관주모형.STM조재결혈재관주전10 min개시이5V、2 ms、1 Hz강도지속전자격경부좌측미주신경20 Bin.재관주120 min후장매조안수궤수자표법분위2부빈,일부빈취좌심실심기조직측정심기경사면적;일부분취좌심실결혈구심기조직성공제취RNA후,획득심기기인표체보.운용GeneSpring연건사선출량조간차이표체현저적기인병진행분석,대기중구유대표의의적기인진행실시정량역전록다취매련반응(Real-time PCR)험증.결과 (1)STM조심기경사구면적교I/R조명현축소(P＜0.05).(2)기인표체보사선득도186개차이표체현저적기인혹탐침,기인본체분석(GO)현시주요영향료3류기인적개변,상조대표기인여심기보호유관;하조대표기인여염증유관.(3)Real-time PCR결과화기인심편표체적결과상부.결론 전자격미주신경대결혈재관주심기구유보호작용,기심기기인표체보발생료흔대적변화,가능통과흥강담감능항염통로래감경화억제심기결혈재관주손상.
Objectives To investigate the effect of electric vagal stimulation on the gene expression of injured myocardium from ischemia/reperfusion rat model and explore the involved molecular mechanism.Methods Twenty Sprague Dawley male rats were randomly selected and divided into 2 groups evenly:ischemia/reperfusion group (I/R group),and vagus nerve stimulation group (STM group).The left anterior descending coronary artery (LAD) was ligated and subjected to 30 min of myocardial ischemia followed by 2 h of reperfusion.In addition,10 min before reperfusion,left cervical vagus nerve of STM group was subjected to electronic stimulation at 5 V,2 ms and 1 Hz for 20 min.After 120 min of reperfusion,every group was randomly divided into two parts.One part that myocardium was collected from left ventricle was applied to determine the area of myocardial infarction.The RNA isolated from another part of the ischemic myocardium collected from left ventricle was hybridized to get gene expression profiles and the quality of hybridized RNA from both I/R and STM group was assessed and analyzed.GeneSpring software was applied to screen out the genes,which show significant difference between groups I/R and STM.Real-time polymerase chain reaction (RT-PCR) was applied to analyze the expression of important genes.Results (1)The area of myocardial infarction of STM (25.5 ± 3.9) ％ was significant reduced relative to L/R group (45.5 ± 4.8) ％ (P ＜ 0.05).(2)The expression levels of 186 genes were changed significantly,and analyzed by Gene ontology (GO) software,there were 3 kinds of genes were affected.The upregulated genes were reported to show protective effect on myocardium.The downregulated gene was relative to inflammation.(3)The RT-PCR result confirmed the genechip assay.Conclusions Electric vagal stimulation can reduce myocardial I/R injury in rats.Significant change of the gene expression was detected between groups I/R and STM.The results suggest that activation of cholinergic anti-inflammatory pathway be involved in the mechanism.