长链非编码RNA-NEAT1参与正畸压力区牙周膜干细胞自噬的研究

Orthodontic treatment induces periodontal tissue remodeling through mechanical system. Periodontal ligament stem cells play an important role in orthodontic tooth movement. It is necessary to understand their biological effects during orthodontic treatment. Autophagy has been seen as an adaptive response to stress. However, the autophagic activity of periodontal ligament stem cells under orthodontic force and the roles of long non-coding RNA (lncRNA) in this process are still not clear. In this study, we found that the expression of lncRNA NEAT1 and the cell autophagic activity were modulated and significantly correlated under mechanical force stimulation in periodontal ligament stem cells. We intended to determine whether NEAT1 mediates autophagic activity of periodontal ligament stem cells under orthodontic stress force. We try to investigate the expression of lncRNA NEAT1 and autophagic activity of periodontal ligament cells under orthodontic stress force in vivo and in vitro; and determine whether NEAT1 knockdown regulates the autophagic activity of periodontal ligament stem cells and orthodontic tooth movement; and finally explore the molecular mechanism whether NEAT1 serves as a ceRNA to active AKT signaling pathway by sponging miR-let7a and miR-320b. This project will be helpful to clarify the role and mechanism of NEAT1 and autophagy in orthodontic remodeling of periodontal tissues.

正畸治疗通过力学系统对牙齿及牙周组织产生组织改建，牙周膜干细胞在正畸牙移动中发挥重要作用，因此，了解正畸力下其生物学行为有重要的意义。自噬是细胞的一种应激反应，在正畸压力下牙周膜干细胞的自噬活性尚不明确，长链非编码RNA（lncRNA）在其中的作用也尚不明确。我们在预实验中发现牙周膜干细胞在机械压力刺激下lncRNA NEAT1及细胞自噬活性明显变化，并呈显著正相关。本项目拟明确NEAT1是否参与正畸压力区牙周膜干细胞自噬调控。为明确这一假设，拟通过体内、体外实验明确正畸力对牙周膜干细胞自噬活性的改变及NEAT1表达的变化；并在干扰NEAT1后检测牙周膜的细胞自噬活性以及牙齿移动情况；通过分子生物学手段研究NEAT1是否通过ceRNA机制吸附miR-let7a及miR-320b活化AKT通路调控细胞自噬。通过以上研究明确NEAT1及细胞自噬在正畸牙周组织改建中的作用及机制。

正畸治疗通过力学系统对牙齿及牙周组织产生组织改建，牙周膜细胞在正畸牙移动中发挥重要作用，因此，了解正畸力下其生物学行为有重要的意义。自噬是细胞的一种应激反应，在正畸压力下牙周膜细胞的自噬活性尚不明确，长链非编码RNA(lncRNA)在其中的作用也尚不明确。本研究明确了lncRNA-FER1L4参与正畸压力区牙周膜细胞自噬调控。通过体内、体外研究模型我们发现在正畸机械压力下牙周膜细胞自噬活性增强，同时长链lncRNA FER1L4的表达显著上调。而干扰FER1L4表达影响了牙周膜细胞的自噬活性并逆转压力下牙周膜细胞的自噬反应。通过分子生物学实验我们揭示FER1L4通过抑制AKT磷酸化，随后增加转录因子FOXO3的核转位，从而介导压应力下的自噬级联反应。FER1L4还可通过吸附miR-874调控VEGFA的表达。通过以上研究我们明确FER1L4及细胞自噬在正畸牙周组织改建中的作用及机制，有利于更好地理解正畸牙移动的机制并进行治疗。