Prevotella intermedia stimulates tissue-type plasminogen activator and plasminogen activator inhibitor-2 expression via multiple signaling pathways in human periodontal ligament cells
Prevotella intermedia stimulates tissue-type plasminogen activator and plasminogen activator inhibitor-2 expression via multiple signaling pathways in human periodontal ligament cells
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DOI：
10.1111/j.1574-695x.2011.00789.x
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发表时间：
2011-06-01
影响因子：
--
通讯作者：
Shu, Lei
中科院分区：
文献类型：
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作者：
Guan, Su-Min;He, Jian-Jun;Shu, Lei
中普氏菌是一种重要的牙周病原菌，可引起多种炎症和免疫反应。在本研究中，我们研究了间质假单胞菌对人牙周韧带（hPDL）细胞纤溶酶原系统的影响，并探讨了其中的信号通路。利用半定量反转录(RT)- pcr和实时定量RT- qpcr，研究人员证实了pif - medium诱导hPDL细胞中组织型纤溶酶原激活物（tPA）和纤溶酶原激活物抑制剂(PAI)-2的表达呈浓度依赖性和时间依赖性，但对尿激酶型纤溶酶原激活物和PAI- 1mrna的表达没有影响。酶联免疫吸附试验证实，普雷沃氏菌中间刺激也能促进tPA蛋白的分泌。Western blot结果显示，P. intermedia处理增加了细胞外信号调节激酶（ERK）、c-Jun n -末端激酶（JNK）和p38激酶（p38）的磷酸化。ERK、JNK和蛋白激酶C抑制剂显著减弱了P. intermedia诱导的tPA和PAI-2的表达。此外，p38和磷脂酰肌醇3-激酶抑制剂显著降低PAI-2的表达，而对tPA的表达没有或几乎没有抑制。与此相反，抑制蛋白激酶A，大大增强了中间芽孢杆菌对tPA和PAI-2表达的上调作用。我们的研究结果表明，中间芽孢杆菌可能通过多种信号通路上调hPDL细胞中tPA和PAI-2的表达，从而促进牙周组织的破坏。
Prevotella intermedia is an important periodontal pathogen that induces various inflammatory and immune responses. In this study, we investigated the effects of P. intermedia on the plasminogen system in human periodontal ligament (hPDL) cells and explored the signaling pathways involved. Using semi-quantitative reverse transcription (RT)-PCR and quantitative real-time RT-qPCR, we demonstrated that P. intermedia challenge increased tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAI)-2 expression in a concentration- and time-dependent manner, but exerted no influence on urokinase-type plasminogen activator and PAI-1mRNA expression in hPDL cells. Prevotella intermedia stimulation also enhanced tPA protein secretion as confirmed by enzyme-linked immunosorbent assay. Western blot results revealed that P. intermedia treatment increased phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 kinase (p38). ERK, JNK and protein kinase C inhibitors significantly attenuated the P. intermedia-induced tPA and PAI-2 expression. Furthermore, p38 and phosphatidylinositol 3-kinase inhibitors markedly decreased PAI-2 expression, whereas they showed no or little inhibition on tPA expression. In contrast, inhibition of protein kinase A greatly enhanced the upregulatory effect of P. intermedia on tPA and PAI-2 expression. Our results suggest that P. intermedia may contribute to periodontal tissue destruction by upregulating tPA and PAI-2 expression in hPDL cells via multiple signaling pathways.