miR-615-3p的抑制通过PVT1介导的线粒体调节增强了肩胛骨中的牙本质形成。

Inhibition of miR-615-3p enhances dentinogenesis in scap via PVT1-mediated mitochondrial regulation.

作者信息

Yang Haoqing, Huang Yishu, Song Jiaxin, Han Xiao, Yuan Fengning, Fan Zhipeng

机构信息

Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, No. 4 Tiantanxili, Dongcheng District, Beijing, China.

Outpatient Department of Oral and Maxillofacial Surgery, School of Stomatology, Capital Medical University, Beijing, China.

出版信息

Stem Cell Res Ther. 2025 Jul 31;16(1):416. doi: 10.1186/s13287-025-04528-7.

DOI:10.1186/s13287-025-04528-7

PMID:40745571

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12315441/

Abstract

BACKGROUND

Mesenchymal stem cells (MSCs) are critical for dental tissue regeneration, yet their differentiation potential is tightly regulated by microRNAs (miRNAs). This study aimed to investigate the role of miR-615-3p in regulating odontogenic differentiation in stem cells from the apical papilla (SCAPs), offering insights into potential applications for enhancing dental tissue regeneration and repair.

METHODS

Quantitative PCR (qPCR), Western blot analysis, alkaline phosphatase (ALP) activity assay, and Alizarin Red staining (ARS) were performed to assess odontogenic differentiation following miR-615-3p modulation in SCAPs. Mitochondrial function was evaluated by measuring reactive oxygen species (ROS) levels, membrane potential, and respiratory activity. In vivo, SCAPs with miR-615-3p modulation were transplanted into rabbit extraction sockets to examine dentin-like tissue formation.

RESULTS

miR-615-3p was significantly downregulated in SCAPs compared to umbilical cord mesenchymal stem cells (WJCMSCs) and further decreased during mineralization induction, suggesting its negative regulatory role in odontogenic differentiation. Inhibition of miR-615-3p enhanced ALP activity, mineralization, and odontogenic marker expression both in vitro and in vivo. Proteomic analysis revealed that miR-615-3p inhibition improved mitochondrial function by reducing ROS levels and increasing mitochondrial function. Further Competing Endogenous RNA Sequencing(ceRNA-seq) analysis identified PVT1 as a downstream target of miR-615-3p. PVT1 overexpression promoted odontogenic differentiation and mitochondrial homeostasis, while its knockdown impaired these processes. Collectively, the miR-615-3p/PVT1 axis emerged as a critical regulator of dentinogenesis through mitochondrial modulation.

CONCLUSIONS

Inhibiting miR-615-3p fosters dentinogenesis through PVT1-mediated mitochondrial regulation in SCAPs. These findings highlight the miR-615-3p/PVT1 axis as a promising target for enhancing dentin tissue engineering applications.

摘要

背景

间充质干细胞（MSCs）对牙组织再生至关重要，但其分化潜能受到微小RNA（miRNAs）的严格调控。本研究旨在探讨miR-615-3p在调节根尖乳头干细胞（SCAPs）成牙分化中的作用，为增强牙组织再生和修复的潜在应用提供见解。

方法

进行定量PCR（qPCR）、蛋白质免疫印迹分析、碱性磷酸酶（ALP）活性测定和茜素红染色（ARS），以评估SCAPs中miR-615-3p调节后的成牙分化情况。通过测量活性氧（ROS）水平、膜电位和呼吸活性来评估线粒体功能。在体内，将经miR-615-3p调节的SCAPs移植到兔拔牙窝中，以检查牙本质样组织的形成。

结果

与脐带间充质干细胞（WJCMSCs）相比，SCAPs中miR-615-3p显著下调，且在矿化诱导过程中进一步降低，表明其在成牙分化中起负调控作用。抑制miR-615-3p可在体外和体内增强ALP活性、矿化和成牙标记物表达。蛋白质组学分析表明，抑制miR-615-3p可通过降低ROS水平和增加线粒体功能来改善线粒体功能。进一步的竞争性内源RNA测序（ceRNA-seq）分析确定PVT1为miR-615-3p的下游靶点。PVT1过表达促进成牙分化和线粒体稳态，而其敲低则损害这些过程。总的来说，miR-615-3p/PVT1轴通过线粒体调节成为牙本质形成的关键调节因子。