电针预处理对心肌缺血再灌注损伤小鼠心肌长链非编码RNA差异表达的影响  

Effect of electroacupuncture pretreatment on differential expression profiles of myocardial long non-coding RNA and mRNA in mice with myocardial ischemia-reperfusion injury

作　　者：徐建俊[1] 张亮[1] 葛文[1] 雍玥 谢晨龙 冯吉杰 王珂 周嘉 XU Jian-jun;ZHANG Liang;GE Wen;YONG Yue;XIE Chen-long;FENG Ji-jie;WANG Ke;ZHOU Jia(Shuguang Hospital Affiliated to the Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海中医药大学附属岳阳中西医结合医院,上海200437

出　　处：《针刺研究》2020年第5期389-395,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81503440、81573796、81603702);上海市进一步加快中医药事业发展三年行动计划[No.ZY(2018-2020)-CCCX-2004-04];2018年上海市卫生计生系统百人计划(No.2018BR24);上海市中西医临床协作试点建设项目[No.ZY(2018-2020)-FWTX-1011]。

摘　　要：目的: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与LncRNA的相关性。方法:将C57BL/6小鼠随机分为假手术组、模型组和电针组,每组4只。采用左前降支冠状动脉结扎法制备MIRI小鼠模型,假手术组仅穿线不结扎。电针组于造模前电针干预双侧"内关"穴30 min。利用高通量LncRNA芯片技术检测各组小鼠心肌组织中差异表达的LncRNA和mRNA,筛选出与电针预处理改善MIRI相关的LncRNA和mRNA并进行生物信息学分析。结果:假手术组、模型组和电针组的LncRNA和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1 693条、mRNA共2 858条。与模型组比较,电针组差异表达的LncRNA共3 859条,mRNA共1 343条。通过Venn交集分析,筛选出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。通过LncRNA-mRNA共表达分析和基因本体富集分析提示,针刺治疗相关的LncRNA可能是通过调控心肌细胞创伤后应激和修复功能参与对MIRI的干预。同时,筛选出来与针刺治疗相关的mRNA其编码的蛋白功能主要涉及细胞创伤后应激和炎性反应调节。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参与了电针预防MIRI的过程,为后续深入研究电针治疗MIRI的作用机制提供了方向和分子依据。Objective To observe the effect of electroacupuncture(EA) pretreatment at "Neiguan"(PC6) on expression profiles of myocardial long non-coding RNAs(LncRNAs) and mRNAs in myocardial ischemia-reperfusion injury(MIRI) mice, so as to explore its mechanisms underlying prevention of MIRI via regulating LncRNA expression.Methods C57 BL/6 mice were randomly divided into sham, model, and EA groups(n =4 in each group). The MIRI model was established by occlusion of the anterior descending branch(ADB) of the left coronary artery for 30 min, followed by reperfusion for 24 h. In the sham group, the ADB was only threaded beneath the artery without ligation. EA was applied to bilateral PC6 for 30 min prior to ischemia induction. Surgery was performed within 30 min at the end of EA stimulation. The expression profiles of differentially expressed LncRNAs and mRNAs in the left ventricular myocardium were analyzed by using LncRNA microarray. Results There was a significant diffe-rence in the expression pattern of LncRNAs and mRNAs among the sham, model and EA groups. A total of 1 693 LncRNAs and 2 858 mRNAs between the model and sham groups, and 3 859 LncRNAs and 1 343 mRNAs between the EA and model groups were identified to be differentially expressed candidates. According to Venn intersection analysis, LncRNAs with opposite regulative orientations in the model and EA groups were screened and defined as EA-related LncRNAs. LncRNA-mRNA co-expression analysis and Gene Ontology enrichment analysis of the EA-related LncRNAs predicted their roles to regulating post-traumatic stress and repairing of myocardial cells. Meanwhile, the proteins’ function encoded by EA-related mRNAs mainly involved post-traumatic stress and inflammatory regulation.Conclusion EA pretreatment at PC6 acupoint can produce extensive regulation on myocardial LncRNAs and mRNAs in MIRI mice, suggesting an involvement of LncRNAs in EA pretreatment induced improvement of MIRI. These results may provide direction and molecular basis for subsequent in-depth studies to

关 键 词：电针 内关 长链非编码RNA 心肌缺血再灌注损伤 

分 类 号：R245.97[医药卫生—针灸推拿学]