State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, 710032, People's Republic of China.
Department of Stomatology Northwest Women's and Children's Hospital, Xi'an, Shaanxi, People's Republic of China.
Stem Cell Res Ther. 2022 Jul 15;13(1):305. doi: 10.1186/s13287-022-02992-z.
High glucose-induced damage to the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) has long been a challenge to periodontal regeneration for diabetic individuals. Metformin is an anti-hyperglycemic drug that exhibits abundant biological activities associated with cell metabolism and downstream tissue regeneration. However, how metformin combats damage to PDLSC osteogenic differentiation under high glucose and the underlying mechanisms remain unknown.
Osteogenic differentiation of PDLSCs was assessed by alkaline phosphatase (ALP) staining, ALP activity, Alizarin Red staining and quantitative assay, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. RNA-seq analysis was performed to screen target genes of metformin, and the effects of target genes were confirmed using lentivirus transfection. Western blot analysis was also used to detect the protein level of underlying signaling pathways.
We found that osteogenic differentiation of PDLSCs under high glucose was decreased, and metformin addition enhanced this capacity of differentiation. Furthermore, the results of RNA-seq analysis showed that natriuretic peptide receptor 3 (NPR3) was upregulated in PDLSCs under high glucose and downregulated after metformin addition. When the underlying pathways involved were investigated, we found that upregulation of NPR3 can compromise the metformin-enhanced PDLSC osteogenic differentiation and activate the MAPK pathway (especially the p38 MAPK and Erk1/2 pathway), and that inhibition of the NPR3-mediated p38 MAPK or Erk1/2 pathway enhanced the osteogenic differentiation of PDLSCs under high glucose.
The present study suggests that metformin may enhance the osteogenic differentiation of PDLSCs under high glucose via downregulation of NPR3 and inhibition of its downstream MAPK pathway. This is the first report identifying the involvement of NPR3-mediated MAPK pathway in the metformin-enhanced osteogenic differentiation, indicating that NPR3 antagonists, such as metformin, may be feasible therapeutics for periodontal tissue regeneration in diabetic individuals.
高糖诱导的人牙周膜干细胞（PDLSCs）成骨分化损伤一直是糖尿病患者牙周再生的挑战。二甲双胍是一种抗高血糖药物，具有丰富的与细胞代谢和下游组织再生相关的生物学活性。然而，二甲双胍如何对抗高糖条件下 PDLSC 成骨分化损伤及其潜在机制尚不清楚。
通过碱性磷酸酶（ALP）染色、ALP 活性、茜素红染色和定量分析、实时定量聚合酶链反应（qRT-PCR）和 Western blot 分析评估 PDLSCs 的成骨分化。进行 RNA-seq 分析以筛选二甲双胍的靶基因，并使用慢病毒转染确认靶基因的作用。Western blot 分析也用于检测潜在信号通路的蛋白水平。
我们发现高糖条件下 PDLSCs 的成骨分化能力下降，而添加二甲双胍增强了这种分化能力。此外，RNA-seq 分析结果表明，高糖条件下 PDLSCs 中钠尿肽受体 3（NPR3）上调，添加二甲双胍后下调。当研究涉及的潜在途径时，我们发现 NPR3 的上调会损害二甲双胍增强的 PDLSC 成骨分化并激活 MAPK 途径（特别是 p38 MAPK 和 Erk1/2 途径），并且抑制 NPR3 介导的 p38 MAPK 或 Erk1/2 途径增强了高糖条件下 PDLSCs 的成骨分化。
本研究表明，二甲双胍可能通过下调 NPR3 及其下游 MAPK 途径增强高糖条件下 PDLSCs 的成骨分化。这是首次报道 NPR3 介导的 MAPK 途径参与二甲双胍增强的成骨分化，表明 NPR3 拮抗剂，如二甲双胍，可能是糖尿病患者牙周组织再生的可行治疗方法。
Zotero 插件
