LncRNA-n285613作为ceRNA参与心肌纤维化调控的功能及机制研究

Cardiac fibrosis is an important pathological factor leading to heart disease, but its molecular mechanism is not fully clarified, and it is lacking of effective interventions. We found that lncRNA-n285613 was significantly increased in fibrotic cardiac tissue. Silencing n28613 was able to inhibit proliferation and collagens generation of cardiac fibroblasts. Bioinformatic analysis revealed that miR-101a and miR-139-5p were predicted targets for n285613. In addition, studies have shown that miR-101a and miR-139-5p respectively regulate the classical pathways of fibrosis, c-fos/TGFβ1 and Wnt/β-catenin. Therefore, we hypothesize that lncRNA-n285613 is able to regulate cardiac fibrosis by targeting miR-101a/c-fos/TGFβ1 and miR-139-5p/Wnt/β-catenin pathways. In the current project, we will employ the genetic manipulated mouse model, cell biology and molecular biology techniques to elucidate the effect and underling mechanism of lncRNA-n285613 in regulating cardiac fibrosis. Thereby it provides the new targets and strategies for the prevention and treatment of cardiac fibrosis.

心肌纤维化是导致心衰等心脏疾病的主要病理因素，其确切分子机制仍未十分清楚，缺少有效干预措施。我们研究发现，lncRNA-n285613在纤维化心肌中表达明显升高，沉默n285613可抑制心肌成纤维细胞的过度增殖及胶原的合成。数据库分析及检测发现，n285613与miR-101a和miR-139-5p具有潜在作用关系，而miR-101a和miR-139-5p可分别调控纤维化的经典通路c-fos/TGFβ1和Wnt/β-catenin。因此，本研究提出假设，lncRNA-n285613可分别通过miR-101a/c-fos/TGFβ1和miR-139-5p/Wnt/β-catenin双重途径调控心肌纤维化。为证实以上假设，本研究将采用转基因动物、细胞生物学及分子生物学等研究手段，对n285613在心肌纤维化中的调控功能及机制进行阐明，为心肌纤维化的研究与防治提供新靶点和新策略。

本研究采用病毒转染、RNA干扰、实时定量PCR及报告基因等技术，分别从在体、离体水平探讨了lncRNAs、miRNAs对心肌纤维化、心肌肥厚及心脏衰老的调控机制。主要发现：1.小鼠心肌纤维化中lncRNA-n285613（2271）表达升高，沉默其表达具有显著的抗心肌纤维化作用；2. m6A甲基转移酶METTL3参与心肌纤维化的调控，lncRNA-2271可通过影响METTL3泛素化调节心肌纤维化；3. lncRNA-CHAR、miR-20b参与心肌肥厚的调控，是心肌肥厚的重要调控分子；4.lncRNA-H19在心肌细胞衰老过程中起到重要调控作用。以上研究发现了心肌纤维化、心肌肥厚及心脏衰老的调控新分子，揭示了心肌纤维化、肥厚及衰老的病理新机制。该发现拓展和加深了人们对lncRNAs、miRNAs在心脏疾病中的调控作用认识。