长链非编码RNA在高血压血管重建中的作用及其力学生物学机制

Long noncoding RNA (lncRNA) are the least well-understood products of transcription from genomes. They do not encode proteins. However, recent explosion in knowledge demonstrating the importance of lncRNA in the regulation of multiple major biological processes impacting on normal life activities and disease have brought these heretofore neglected molecular players to forefront. Although new lncRNA are being discovered at a rapid pace, the contributions of lncRNA to cardiovascular mechanobiology remain largely unknown. In this project, first, the genetic differences map of lncRNAs between spontaneously hypertensive rats (SHR) and wistar kyoto rats (WKY) is created by gene chip. And then the obtained data are clustered and analyzed by using bioinformatics method. In the second step, the role of the screened lncRNA in regulation of proliferation and migration in VSMCs subjected to cyclic mechanical strain by Flexcell-4000 is studied. Finally, the new screened lncRNA is knocked down in SHR by RNAi in vivo, leading to identification of its role in vascular remodeling. With the employment of recent advances in technologies, i.e. gene chip、bioinformatics method、mechanical cell biology and molecular biology, the project will achieve successes in illuminating the role of lncRNA in hypertension-induced vascular remodeling. Further, the accomplishment of the project will provide deep insights into the mechanobiological mechanism and potential targets for hypertension therapy.

长链非编码RNA（lncRNA）是基因组非编码序列的转录产物，其本身不编码蛋白质，但对基因表达具有重要的调控作用，在正常生命活动和疾病发生中发挥重要作用。但是，迄今为止，对它的研究刚刚起步。有关lncRNA的研究尚未涉及心血管力学生物学领域。本项目拟利用基因芯片技术，建立自发性高血压大鼠（SHR）与对照大鼠（WKY）主动脉lncRNA差异表达谱，采用生物信息学方法对差异lncRNA进行聚类和分析；利用细胞体外周期性张应变加载模型，研究筛选到的lncRNA在张应变调控VSMCs增殖和迁移中的作用；最后，回归高血压模型，在体条件下检测筛选到的lncRNA对血压及血管重建的影响。本项目应用基因组学、生物信息学、细胞力学和分子生物学等相关学科理论和实验技术，旨在明确lncRNA在高血压血管重建中的作用及其力学生物学机制，为阐明高血压血管重建的发病机理和药物治疗靶标的研究提供新的力学生物学依据。

本项目按着研究计划，应用基因芯片技术、生物信息学分析方法、细胞生物力学和经典分子生物学研究方法，探讨了长链非编码RNA (long noncoding RNA，lncRNA)在高血压诱导的血管重建中的作用。首先应用基因芯片发现在自发性高血压大鼠（SHR）与对照大鼠胸主动脉（WKY）中，lncRNA和mRNA表达不同。 同WKY相比，SHR中68个lncRNA分子上调，167个lncRNA分子下调。SHR中255个mRNA上调，272个mRNA下调。然后结合PCR验证结果和表达值构建基因共表达网络以及IPA数据库分析，重点关注了lncRNA MRAK048635和XR007793的功能。研究结果揭示MRAK048635和Sirt6在张应变诱导的血管平滑肌细胞（vascular smooth muscle cells，VSMCs）分化中可能起重要作用。而且XR007793与VSMCs增殖功能变化密切相关，其在静态条件下可能通过调控Stat2、Lmo2、Irf7的表达变化参与VSMCs增殖，在高张应变条件下可能通过调控Cdca8的表达变化参与高周期性张应变诱导的VSMCs增殖变化过程。以上研究结果提示lncRNA在应力信号转导网络中具有重要作用，丰富了高血压条件下血管重建的力学生物学机制，并为寻找高血压药物治疗靶标提供了新思路。