电针“内关”穴预处理对心肌缺血再灌注损伤保护的lncRNA靶基因筛选及其生物信息学挖掘

Myocardial ischemia-reperfusion injury (IRI) in cardiopulmonary bypass cardiac surgery remains the most common and the key issue to be solved. LncRNA, participating in a variety of important regulatory process, such as chromatin modification, transcriptional activation, transcription interference, nuclear transport, is relevant to physiological and pathological processes of cardiac function, myocardial ischemia, and etc. From previous studies on acupuncture anesthesia, we confirmed that acupuncture anesthesia has effects on heart protection. Based on the above clinical research and recent literature research, as well as on the relationships between the circulating lncRNA and acupuncture anesthesia, this study ①analyzes time-effect relationship of transcutaneous acupoint electrical stimulation (TAES) on myocardial IRI protection, with a focus on protection of cardiac function; ②observes the fluctuations of TAES-pretreated lncRNA using the IRI model in rats and discover the corresponding target gene; ③inspects the myocardial lncRNA target gene expression in the IRI model in rats within the most effective time, and identify the change of specific lncRNA in myocardium and peripheral circulation. Finally, this study will reveal the acupuncture– lncRNA – target protein network regulatory mechanism, which will provide scientific basis for clinical application and reveal the material base of acupuncture in heart operation.

心脏手术体外循环过程中出现的心肌缺血-再灌注损伤是最常见而有待于解决的关键问题。lncRNA参与染色质修饰，转录激活，转录干扰，核内运输等多种重要的调控过程，与心脏功能、心肌缺血等多种生理病理过程相关。通过前期针刺的研究，我们发现针刺在在手术中具有心脏保护作用。基于上述临床研究基础和最新文献检索，围绕循环lncRNA与针刺效应之间的关系，本课题①分析电针内关在心肌缺血再灌注损伤保护中的时间效用分析，重点观察心功能保护时间效应关系；②利用大鼠缺血再灌注模型，观察电针内关预处理后的lncRNA的变化，及发现相应的靶基因；③在最佳效应时间内，检测大鼠缺血再灌注模型的心肌lncRNA靶基因表达水平，明确特异lncRNA在心肌中和外周循环的变化情况。最终揭示针刺 “效应-lncRNA-靶基因”多靶点多层次的综合调节机制，为临床在心脏手术中推广和应用针刺解决关键科学基础问题和效应物质基础。

心脏手术体外循环过程中出现的心肌缺血-再灌注损伤是最常见而有待于解决的关键问题。lncRNA参与染色质修饰，转录激活，转录干扰，核内运输等多种重要的调控过程，与心脏功能、心肌缺血等多种生理病理过程相关。通过前期针刺的研究，我们发现针刺在在手术中具有心脏保护作用。观察电针“内关”预处理对心肌缺血再灌注损伤（MIRI）小鼠心肌长链非编码RNA（LncRNA）和mRNA表达的影响，探讨电针预处理抗MIRI的作用与lncRNA的相关性。方法：C57BL/6小鼠随机分为假手术组、模型组和电针组，每组8只。采用左前降支冠状动脉结扎术，缺血30 min再灌注24 h制备小鼠MIRI，假手术组仅穿线不结扎。电针组选取双侧“内关”穴，于造模前电针干预30 min，针刺刺激结束后30 min内对小鼠进行手术造模。利用高通量lncRNA芯片技术检测模型组和电针组小鼠心肌组织中差异表达的lncRNA和mRNA，筛选出电针治疗心肌缺血再灌注损伤相关的lncRNA和mRNA。结果：假手术组、模型组和电针组之间的lncRNA-mRNA的整体表达特征存在明显差异。与假手术组比较，模型组差异表达的lncRNA共1725条、mRNA共2858条。与模型组比较，电针组差异表达的lncRNA共3867条，mRNA共1343条。对差异mRNA进行生物信息学分析，结果显示模型组差异mRNA主要涉及免疫炎症反应相关通路，电针调节的差异mRNA主要涉及神经-免疫调节相关通路。通过韦恩分析，筛选出在491条在模型组和电针组调节方向相反的lncRNA，定义为针刺治疗相关的LncRNA。结论：电针“内关”预处理能对MIRI小鼠心肌lncRNA和mRNA产生广泛的调节，提示lncRNA参与了电针治疗MIRI过程，为后续深入研究揭示电针治疗MIRI效应提供了方向和分子依据。