长链非编码RNA RP5-833A20.1对NFIA的调控及其在胆固醇平衡和动脉粥样硬化发生发展中的作用

Long non-coding RNA (lncRNA) plays a crucial role in regulation of gene expression and is associated with many diseases. However, the effect of lncRNA on cardiovascular diseases is still unknown. Microarray analysis revealed that expression of lncRNA RP5-833A20.1 was up-regulated while expression of NFIA was down-regulated in THP-1 macrophage-derived foam cells. Furthermore, RP5-833A20.1 reduced macrophage cholesterol efflux through inhibiting NFIA expression. Thus, we hypothesized that atherosclerosis development can be affected by RP5-833A20.1-NFIA through regulating gene and protein expression involved in maintaining cholesterol homeostasis. In this project, we aim to explore the effects of RP5-833A20.1-NFIA on gene and protein expression involved in cholesterol metabolism and on atherosclerosis development by enhancing expression of RP5-833A20.1 and/or NFIA. Therefore, our study may provide new scientific evidence to establish RP5-833A20.1- NFIA as a new target for regulation of cholesterol homeostasis and prevention of atherosclerosis.

长链非编码RNA(lncRNA)在基因调控中起关键作用并与众多疾病相关，但其与心血管疾病的研究才刚起步。我们首次用lncRNA和mRNA表达谱芯片进行THP-1巨噬细胞和THP-1巨噬细胞源性泡沫细胞中lncRNA和mRNA差异筛选，发现THP-1巨噬细胞源性泡沫细胞中lncRNA RP5-833A20.1上调而NFIA下调。进一步实验表明NFIA促进巨噬细胞胆固醇流出而RP5-833A20.1下调NFIA表达并抑制巨噬细胞胆固醇流出。因此我们首次提出RP5-833A20.1-NFIA可能通过调节维持胆固醇平衡相关基因和蛋白表达来影响As形成。本项目拟通过增强RP5-833A20.1和/或NFIA表达，观察RP5-833A20.1-NFIA对维持胆固醇平衡关键基因和蛋白表达的影响及其对As形成的影响，为确立RP5-833A20.1-NFIA作为调控胆固醇平衡和防治As新靶点提供科学依据。

长链非编码RNA(lncRNA)在基因调控中起关键作用并与众多疾病相关，但其与心血管疾病的研究才刚起步。我们首次用lncRNA和mRNA表达谱芯片进行THP-1巨噬细胞和THP-1巨噬细胞源性泡沫细胞中lncRNA和mRNA差异筛选，发现THP-1巨噬细胞源性泡沫细胞中lncRNA RP5-833A20.1上调而NFIA下调。分别构建RP5-833A20.1的慢病毒过表达载体、NFIA的慢病毒过表达载体并将载体分别转染THP-1巨噬细胞，观察其对THP-1巨噬细胞胆固醇摄取和流出的影响。NFIA 的慢病毒过表达载体尾静脉注射apoE-/-小鼠体内，使其在小鼠体内稳定过表达NFIA，观察过表达NFIA对小鼠体内ABCA1、ABCG1、SR-B1、LDLR、NPC1L1和ABCG5/ABCG8 等表达的影响，观察小鼠体内胆固醇逆向转运效率的变化，观察小鼠腹腔巨噬细胞胆固醇流出的变化，同时观察其对小鼠动脉粥样硬化病变的影响。本研究发现Ox-LDL和Ac-LDL可上调RP5-833A20.1表达，抑制NFIA表达。RP5-833A20.1通过上调has-miR-382-5P表达进而抑制NFIA表达。过表达RP5-833A20.1或者过表达has-miR-382-5P均可促进Ox-LDL的摄取，增加细胞内胆固醇含量，减少胆固醇流出，促进炎症因子表达，过表达NFIA则作用相反。NFIA促进小鼠体内胆固醇逆向转运，减少循环炎症因子表达。过表达NFIA抑制小鼠体内动脉粥样硬化形成。本研究发现RP5-833A20.1/has-miR-382-5P/NFIA通路调控细胞内胆固醇平衡及炎性因子的表达，为确立RP5-833A20.1/has-miR-382-5P/NFIA作为调控胆固醇平衡和动脉粥样硬化防治靶点提供科学依据。