Zhou Min, Li Zhuo-quan, Wang Zuo-lin
Shanghai Engineering Research Center of Tooth Restoration and Regeneration, School of Stomatology, Tongji University, Shanghai 200072, China.
Shanghai East Hospital, The Institute for Biomedical Engineering and Nano Science, Tongji University School of Medicine, Tongji University, Shanghai 200092, China.
Acta Pharmacol Sin. 2015 Nov;36(11):1388-94. doi: 10.1038/aps.2015.77. Epub 2015 Oct 26.
S100A4, also known as fibroblast-specific protein 1 or metastasin 1, is not only highly expressed in growth-stimulated cultured cells and metastatic tumor cells, but also in the periodontal ligament. The aim of this study was to investigate the roles of S100A4 in the pathogenesis of periodontitis and its regulatory mechanisms in inflammatory milieu.
Experimental periodontitis was induced in rats by submarginal silk ligatures. TRAP activity and S100A4 expression in periodontal ligaments were examined using immunohistochemistry and immunofluorescence methods. IL-1β-treated human periodontal ligament cells (hPDLCs) were used as in vitro model of experimental periodontitis. S100A4 mRNA and protein were assessed using qRT-PCR and Western blot, respectively. hPDLCs were transfected with either S100A4 overexpression plasmids or shRNAs plasmids. The mineralization in hPDLCs was evaluated with a 12-d osteogenic induction assay, and the expression of ALP, OCN, MMP-2 and MMP-13 was analyzed by qRT-PCR.
In the periodontal ligaments of rats with experimental periodontitis, TRAP activity and S100A4 protein staining were considerably more intense compared with those in the control rats. Treatment of hPDLCs with IL-1β (10, 50 and 100 ng/mL) dose-dependently increased the mRNA and protein levels of S100A4. Transfection with shRNAs markedly increased mineralized nodule formation and the osteogenic-related markers ALP and OCN levels in hPDLCs, whereas the overexpression of S100A4 significantly reduced mineralized nodule formation, and increased the matrix degradation enzymes MMP-2 and MMP-13 levels in hPDLCs.
S100A4 is upregulated in the experimental rat periodontitis and in IL-1β-treated hPDLCs, where S100A4 suppresses osteogenic differentiation and enhances matrix degradation. Thus, S100A4 is a potential target for the treatment of periodontitis.
S100A4，也被称为成纤维细胞特异性蛋白1或转移相关蛋白1，不仅在生长刺激的培养细胞和转移性肿瘤细胞中高表达，在牙周韧带中也有高表达。本研究旨在探讨S100A4在牙周炎发病机制中的作用及其在炎症环境中的调控机制。
通过龈下丝线结扎诱导大鼠实验性牙周炎。采用免疫组织化学和免疫荧光方法检测牙周韧带中的抗酒石酸酸性磷酸酶（TRAP）活性和S100A4表达。用白细胞介素-1β（IL-1β）处理的人牙周膜细胞（hPDLCs）作为实验性牙周炎的体外模型。分别用qRT-PCR和蛋白质免疫印迹法评估S100A4的mRNA和蛋白质水平。用S100A4过表达质粒或短发夹RNA（shRNAs）质粒转染hPDLCs。通过12天的成骨诱导试验评估hPDLCs中的矿化情况，并用qRT-PCR分析碱性磷酸酶（ALP）、骨钙素（OCN）、基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-13（MMP-13）的表达。
在实验性牙周炎大鼠的牙周韧带中，与对照大鼠相比，TRAP活性和S100A4蛋白染色明显更强。用IL-1β（10、50和100 ng/mL）处理hPDLCs可剂量依赖性地增加S100A4的mRNA和蛋白质水平。用shRNAs转染显著增加了hPDLCs中矿化结节的形成以及成骨相关标志物ALP和OCN的水平，而S100A4的过表达显著减少了矿化结节的形成，并增加了hPDLCs中基质降解酶MMP-2和MMP-13的水平。
S100A4在实验性大鼠牙周炎和IL-1β处理的hPDLCs中上调，其中S100A4抑制成骨分化并增强基质降解。因此，S100A4是治疗牙周炎的一个潜在靶点。
