基于长链非编码RNA介导血管新生途径研究针刺干预脑梗塞的分子机制

Angiogenesis is an important link in the pathogenesis of stroke,when induction of angiogenesis has become one of the important strategies to treat ischemic stroke.Previous studies have confirmed the exact role of acupuncture in promoting angiogenesis and establishing effective collateral circulation and in the acute phase in rats of cerebral infarction,but the regulatory mechanism of acupuncture is still unclear.LncRNA are involved in the regulation of angiogenesis and play an important role in the pathophysiology of angiogenesis.Foreign studies have confirmed that the expression level of lncRNA changes significantly after cerebral infarction,However,there have been few reports on the relationship between lncRNA and postinfarct angiogenesis and the effect of acupuncture on its expression.Therefore,we propose that there is a difference in gene regulation between acupuncture and the opening and formation of cerebrovascular collateral circulation in the ischemic state,and that acupuncture can mediate the angiogenesis after cerebral infarction by regulating lncRNA.In order to verify this hypothesis,this project intends to apply microarray technology,bioinformatics technology,immunofluorescence double labeling,Western Blotting,qRT-PCR and other technologies to confirm the above hypothesis from the molecular,tissue and overall animal levels,which will reveal the effects and regulatory mechanism of acupuncture on cerebral infarction from the gene level,and provide more scientific experimental basis for the clinical promotion and application of acupuncture on cerebral infarction.

血管新生是脑卒中发病过程中的重要环节，诱导血管生成已成为目前治疗缺血性脑卒中的重要策略之一。前期研究证实了针刺在促进血管新生建立脑梗塞大鼠有效侧支循环的确切作用，但关于针刺作用的调控机制仍不清楚。长链非编码RNA参与血管生成过程，并在血管新生中发挥重要作用。国外研究证实脑梗塞后lncRNA表达水平发生显著变化，但lncRNA与脑梗塞后血管新生的关系及针刺对其表达的影响，目前鲜有报道。因此，我们提出针刺对缺血状态下脑血管侧支循环的开放和形成在基因调控上存在差异，针刺通过调控lncRNA介导脑梗塞后血管新生的假说。为了验证这一假说，本项目拟应用微阵列技术、生物信息学技术、免疫荧光双标记、Western Blotting、qRT-PCR等技术从分子、组织和整体动物等水平多方面证实上述假设。从基因水平揭示针刺治疗脑梗塞的效应及调控机制，为针刺治疗脑梗塞的的临床推广应用提供更多的科学实验依据。

血管新生是脑卒中发病过程中的重要环节，诱导血管生成已成为目前治疗缺血性脑卒中的重要策略之一。前期研究证实了针刺在促进血管新生建立脑梗塞大鼠有效侧支循环的确切作用，但关于针刺作用的调控机制仍不清楚。长链非编码RNA参与血管生成过程，并在血管新生中发挥重要作用。国外研究证实脑梗塞后lncRNA表达水平发生显著变化，但lncRNA与脑梗塞后血管新生的关系及针刺对其表达的影响，目前鲜有报道。本项目应用微阵列技术、生物信息学技术、免疫荧光双标记、Western Blotting、qRT-PCR等技术从分子、组织和整体动物等水平多方面获得：MCAO模型构建后，大量lncRNAs发生上下调改变，电针对lncRNAs的表达存在促进或抑制的双向调节作用，并且通过介导lncRNA调控其下游血管新生关键信号通路VEGF/Notch相关因子的激活，从而促进内皮细胞的增殖、分化和迁移，诱导血管新生，改善MCAO模型大鼠神经功能缺损症状，初步将“醒脑开窍”针刺法与系统生物学中的转录组学相结合，从基因水平揭示了针刺治疗脑梗塞的效应及调控机制。为针刺治疗脑梗塞的临床推广应用提供了更多的科学实验依据。