长链非编码RNA在同型半胱氨酸诱导大鼠血管平滑肌细胞炎性因子表达中的作用

Homocysteine (Hcy) is an independent risk factor for atherosclerosis. We previously demonstrated that Hcy is able to participate in the progress of atherosclerosis through stimulating inflammatory response in vascular smooth muscle cells (VSMCs). However, the molecular mechanisms of pro-inflammatory and pro-atherosclerotic effects of Hcy remain largely unclear. The present project will further explore the pro-inflammatory mechanisms of Hcy in VSMCs. Long noncoding RNA (lncRNA) participates in modulating biological processes through regulating gene expression at almost all levels. In addition, recent studies revealed that lncRNAs play important roles in the process of inflammation and atherosclerosis. However, the role of lncRNA in the inflammation and atherosclerosis resulting from Hcy is still unknown. We plan to screen the differentially expressed lncRNAs in Hcy treated VSMCs by lncRNA microarray, and further to identify the key lncRNA in Hcy-stimulated inflammation in VSMCs. Then, we will try to explore the possible signal transduction pathways interacted with the key lncRNA and to elucidate the possible mechanisms of how Hcy regulates the key lncRNA in the process based on bioinformatics analysis and traditional experiments including RNAi, Western blot, ChIP and RNA-pull down. This project will give new insights to the epigenetic mechanisms of Hcy’s pro-inflammatory effects and may provide a new target for the treatment of HHcy and HHcy related atherosclerosis.

同型半胱氨酸（Hcy）是动脉粥样硬化的独立危险因素，我们前期研究证实Hcy通过刺激血管平滑肌细胞（VSMCs）炎症参与动脉粥样硬化的发生发展，然而其致炎分子机制未完全明确。lncRNA从多个水平参与基因表达和信号通路调控，并参与炎症和动脉粥样硬化进程。但是，lncRNA在Hcy致炎致动脉粥样硬化中的作用尚不清楚。本课题拟通过芯片筛选在Hcy处理的VSMCs中差异表达的lncRNA，并通过生物信息学方法、qRT-PCR、RNAi等技术确定在Hcy刺激VSMCs炎症中的关键lncRNA并验证其功能，继而结合ChIP、RNA-pull down等手段探讨lncRNA参与调控炎症因子表达炎性信号通路及机制，并初步研究Hcy调控lncRNA的机理。这将为Hcy的致炎致动脉粥样硬化作用提供新的表观遗传学机制，并可能为高同型半胱氨酸血症及相关动脉粥样硬化治疗和药物研发提供新的靶点。

同型半胱氨酸（Hcy）是动脉粥样硬化的独立危险因素，我们前期研究进一步证实Hcy通过刺激血管炎症参与动脉粥样硬化的发生发展，然而其致炎分子机制未完全明确。lncRNA从多个水平参与基因表达和信号通路调控，并参与炎症和动脉粥样硬化进程。但是，lncRNA在Hcy致炎致动脉粥样硬化中的作用尚不清楚。本研究通过RNA-seq发现Hcy刺激VSMCs可引起lncRNA表达谱的改变，这提示lncRNA在Hcy的致病过程中发挥重要作用。根据lncRNA差异表达分析、lncRNA靶向mRNA分析及GO富集分析结果，采用real-time PCR选择验证了部分lncRNA的表达水平。并通过过表达和敲低确证了lncRNA H19的作用。初步明确了lncRNA H19在Hcy刺激VSMCs炎症中的关键作用，并证实lncRNA H19通过调控MAPK-NF-kappaB信号通路参与Hcy刺激VSMCs 炎性因子的表达。明确Hcy可通过影响DNA甲基化调控lncRNA H19的表达。这为Hcy的致炎致动脉粥样硬化作用提供新的表观遗传学机制，并可能为高同型半胱氨酸血症及相关动脉粥样硬化治疗和药物研发提供新的靶点。