炎性环境下lncRNA-Gm10435相关的ceRNA网络参与miR-155-3p/经典Wnt信号通路调控成牙骨质细胞分化的机制研究

Currently, many studies showed that the development and differentiation of cementum had critical role in the regeneration of periodontal tissues. Our recent previous study demonstrated the expression of Wnt in cementum as well as the important role of Osx on cementoblast differentiation by regulating Wnt signaling pathway via DKK1. At the same time, we also found that miR-155-3p could participate in cementoblast differentiation by targeting Kctd1 via regulating Wnt signaling pathway. Besides, bioinformatic prediction found that lncRNA-Gm10435 could competitively bind to miR-155-3p; further pre-test by us also discovered the negative correlation of lncRNA-Gm10435 with TNF-α induced inflammation on time levels. Based on above facts, we put forward the hypothesis that lncRNA-Gm10435-related ceRNA network might be involved in cementoblast differentiation under inflammatory conditions by regulating miR-155-3p/canonical Wnt signaling pathway. Hence, on one hand, we will investigate the effect of lncRNA-Gm10435 on cementoblast differentiation by gain/lose-of-function experiments. On the other hand, we will lucubrate the ceRNA regulatory network under inflammatory conditions in vivo and in vitro. The effects of lncRNA-Gm10435/miR-155-3p/Wnt signaling pathway on the formation and mineralization of cementum may guide a great direction of cementum regeneration.

目前研究认为，牙骨质的发育分化调控在牙周组织再生中起重要作用。在前期研究中，本课题组发现Wnt信号通路参与牙骨质中的发育调控，Osx通过DKK1调控Wnt信号通路在成牙骨质细胞分化中起重要作用；同时发现炎性状态下，miR-155-3p也通过靶向Kctd1，调控Wnt信号通路参与成牙骨质细胞的分化；此外通过生物信息学预测 lncRNA-Gm10435能够与miR-155-3p结合，进一步预实验研究也证实lncRNA-Gm10435与TNF-α炎性诱导时间呈负相关。基于上述研究背景，我们提出炎性环境下lncRNA-Gm10435相关的ceRNA网络参与miR-155-3p/经典Wnt信号通路调控成牙骨质细胞分化的机制研究，为此本课题将深入研究体内外炎性状态下这一调控网络对牙骨质形成和矿化的影响，为牙骨质的发育分化的精准调控及牙周组织的再生提供新的切入点和方向。

研究表明长链非编码RNA和信使RNA在细胞矿化和骨的形成中发挥关键作用。因此，我们想要确定在成牙骨质细胞矿化过程中长链非编码RNA和信使RNA的表达谱、表达模式以及这些差异基因的潜在生物学功能。. 成功诱导成牙骨质细胞矿化后，利用高通量测序技术和生物信息学技术，发现与0天相比较，在矿化第7天时，共有384个差异表达的lncRNA和4255个差异表达的mRNA，在矿化第14天时，共有645个差异表达的lncRNA和4717个差异表达的mRNA，并且这些差异表达的基因可分为6种不同的表达模式。选取感兴趣的lncRNA-Platr3和CXXC5,发现在它们能够分别通过NF-κB信号通路和MAPK信号通路来部分调控牙龈卟啉单胞菌抑制的成牙骨质细胞的矿化。同时，我们发现线粒体的生物发生可通过MAPK信号通路逆转低氧环境对成牙骨质细胞矿化的抑制作用。. 上述实验结果为探究成牙骨质细胞的矿化机制提供了新的理论依据，有利于今后牙周组织再生工程的发展。