Shikonin promotes osteogenic differentiation of periodontal ligament stem cells by modulating the p38 MAPK/ETS transcription factor 1/cementum protein 1 axis in vitro
Zeyu Wu, Chao Shan, Jie Song, Qian Yu, Bing-jie Lian, Jin Zhao
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引用次数: 0
Abstract
Periodontitis is a common disorder that causes tooth loss. Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is crucial for repairing periodontal defects. This study aimed to investigate the underlying mechanism by which Shikonin promotes the osteogenic differentiation of PDLSCs. PDLSCs were isolated from the periodontal ligaments of orthodontically extracted premolars. Flow cytometry was used to characterize the cell surface markers of PDLSCs. PDLSCs were cultured in osteogenic induction medium and treated with Shikonin. Alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining were used to evaluate ALP activity and assess calcium nodule formation, respectively. Protein levels were determined by Western blot analysis. A dual-luciferase reporter assay and chromatin immunoprecipitation analysis confirmed the interaction between the ETS transcription factor 1 (ELK1) and cementum protein 1 (CEMP1) promoter. Shikonin promoted the osteogenic differentiation of PDLSCs (about 7.6 folds for ALP staining and about 6.9 folds for ARS staining with 1 μm Shikonin, p < 0.01). Shikonin-mediated enhancement of osteogenic differentiation was achieved by promoting the phosphorylation of ELK1 through the activation of the p38 MAPK signaling pathway (p < 0.05). Moreover, ELK1 accelerated osteogenic differentiation and transcriptionally activated CEMP1 expression (p < 0.05). Activation of the ELK1/CEMP1 axis contributed to Shikonin-induced osteogenic differentiation of PDLSCs (p < 0.05). Shikonin promotes the osteogenic differentiation of PDLSCs in vitro by modulating the p38 MAPK/ELK1/CEMP1 axis.
牙周炎是一种常见的导致牙齿脱落的疾病。牙周韧带干细胞成骨分化是修复牙周缺损的关键。本研究旨在探讨紫草素促进PDLSCs成骨分化的潜在机制。从正畸拔牙的牙周韧带中分离PDLSCs。采用流式细胞术对PDLSCs的细胞表面标志物进行表征。PDLSCs在成骨诱导培养基中培养，并经紫草素处理。碱性磷酸酶（ALP）和茜素红S （ARS）染色分别评价ALP活性和钙结节形成情况。Western blot检测蛋白水平。双荧光素酶报告试验和染色质免疫沉淀分析证实了ETS转录因子1 （ELK1）和骨水泥蛋白1 （CEMP1）启动子之间的相互作用。紫草素促进PDLSCs成骨分化（1 μm紫草素ALP染色约为7.6倍，ARS染色约为6.9倍），p p p p p
期刊介绍：
The Journal of Cell Communication and Signaling provides a forum for fundamental and translational research. In particular, it publishes papers discussing intercellular and intracellular signaling pathways that are particularly important to understand how cells interact with each other and with the surrounding environment, and how cellular behavior contributes to pathological states. JCCS encourages the submission of research manuscripts, timely reviews and short commentaries discussing recent publications, key developments and controversies.
Research manuscripts can be published under two different sections :
In the Pathology and Translational Research Section (Section Editor Andrew Leask) , manuscripts report original research dealing with celllular aspects of normal and pathological signaling and communication, with a particular interest in translational research.
In the Molecular Signaling Section (Section Editor Satoshi Kubota) manuscripts report original signaling research performed at molecular levels with a particular interest in the functions of intracellular and membrane components involved in cell signaling.
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