Regulation of p53 under hypoxic and inflammatory conditions in periodontium
Regulation of p53 under hypoxic and inflammatory conditions in periodontium
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DOI：
10.1007/s00784-015-1679-x
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发表时间：
2016-09-01
影响因子：
3.4
通讯作者：
Goetz, W.
中科院分区：
文献类型：
--
作者：
Memmert, S.;Goelz, L.;Goetz, W.
不同的研究表明炎症和缺氧会导致 p53 蛋白水平增加。然而，p53 在口腔炎症过程中的意义仍不清楚。因此，本研究的目的是在体外和体内研究缺氧和炎症对人牙周组织 p53 调节的影响。在缺氧和常氧条件下，用牙周病原菌牙龈卟啉单胞菌 (P.g.) 的脂多糖 (LPS) 刺激人初级牙周膜 (PDL) 成纤维细胞 (n = 9)。不同时间点后，测试细胞活力；使用定量 RT-PCR、免疫印迹和免疫荧光测量 p53 基因表达、蛋白质合成和激活。此外，从12名捐赠者身上获取健康和发炎的牙周组织，通过免疫组织化学分析口腔炎症疾病中的p53蛋白。缺氧最初诱导 p53 mRNA 表达和 p53 蛋白水平显着上调。炎症刺激后 p53 的核易位支持了这些发现。缺氧首先增强 p53 水平，但孵育 24 小时后，蛋白质水平下降，同时 PDL 细胞活力提高。免疫组织化学显示p53免疫反应性随着牙周炎症的进展而升高。我们的数据表明p53在PDL细胞稳态中发挥着关键作用，并且似乎在口腔炎症疾病中上调。p53的上调可能促进牙周完整性的破坏。可以讨论与致癌作用的可能关系。
Different studies suggest that inflammation as well as hypoxia leads to an increase of p53 protein levels. However, the implication of p53 during oral inflammatory processes is still unknown. The aim of this study was therefore to investigate the effect of hypoxia and inflammation on p53 regulation in human periodontium in vitro and in vivo.Under hypoxic and normoxic conditions, human primary periodontal ligament (PDL) fibroblasts (n = 9) were stimulated with lipopolysaccharides (LPS) from Porphyromonas gingivalis (P.g.), a periodontal pathogenic bacterium. After different time points, cell viability was tested; p53 gene expression, protein synthesis, and activation were measured using quantitative RT-PCR, immunoblotting, and immunofluorescence. Moreover, healthy and inflamed periodontal tissues were obtained from 12 donors to analyze p53 protein in oral inflammatory diseases by immunohistochemistry.LPS-P.g. and hypoxia initially induced a significant upregulation of p53 mRNA expression and p53 protein levels. Nuclear translocation of p53 after inflammatory stimulation supported these findings. Hypoxia first enhanced p53 levels, but after 24 h of incubation, protein levels decreased, which was accompanied by an improvement of PDL cell viability. Immunohistochemistry revealed an elevation of p53 immunoreactivity in accordance to the progression of periodontal inflammation.Our data indicate that p53 plays a pivotal role in PDL cell homeostasis and seems to be upregulated in oral inflammatory diseases.Upregulation of p53 may promote the destruction of periodontal integrity. A possible relationship with carcinogenesis may be discussed.