Pattamapun Kassara, Tiranathanagul Siriluck, Yongchaitrakul Tussanee, Kuwatanasuchat Jintakorn, Pavasant Prasit
Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
J Periodontal Res. 2003 Apr;38(2):115-21. doi: 10.1034/j.1600-0765.2003.01650.x.
It has been reported that matrix metalloproteinase (MMP) produced by host cells plays a major role in periodontal tissue destruction. In addition, secreted virulence factors from Porphyromonas gingivalis can alter MMP secretion and cause activation in host cells that lead to the tissue degradation. In this study, we examine the effects of P. gingivalis supernatant on matrix metalloproteinase-2 (MMP-2) activation in human periodontal ligament (HPDL) cells. Cultures of HPDL cells were treated with P. gingivalis supernatant for 48 h and the level of MMP-2 activation was monitored by gelatin zymography. The profound activation of MMP-2 was seen only in the treated group. The activation of MMP-2 was inhibited by MMP inhibitors phenanthroline and EDTA, but not serine protease or cysteine protease inhibitors. To study the correlation between the expression of membrane-type-1 matrix metalloproteinase (MT1-MMP) and the activation of MMP-2, the level of MT1-MMP was analyzed. The results from reverse-transcription polymerase chain reaction (RT-PCR) and Western analysis indicated that P. gingivalis supernatant up-regulated the expression of MT1-MMP in both transcription and translation levels within 48 h. These results suggest that P. gingivalis supernatant can activate MMP-2 in HPDL cells and the mechanism of activation may involve the increased amount of MT1-MMP. It is possible that the activation of MMP-2 by P. gingivalis plays a role in the process of chronic periodontitis.
据报道，宿主细胞产生的基质金属蛋白酶（MMP）在牙周组织破坏中起主要作用。此外，牙龈卟啉单胞菌分泌的毒力因子可改变MMP的分泌，并导致宿主细胞中的激活，从而导致组织降解。在本研究中，我们检测了牙龈卟啉单胞菌上清液对人牙周膜（HPDL）细胞中基质金属蛋白酶-2（MMP-2）激活的影响。用牙龈卟啉单胞菌上清液处理HPDL细胞培养物48小时，并通过明胶酶谱法监测MMP-2的激活水平。仅在处理组中观察到MMP-2的显著激活。MMP-2的激活被MMP抑制剂菲咯啉和EDTA抑制，但不被丝氨酸蛋白酶或半胱氨酸蛋白酶抑制剂抑制。为了研究膜型-1基质金属蛋白酶（MT1-MMP）的表达与MMP-2激活之间的相关性，分析了MT1-MMP的水平。逆转录聚合酶链反应（RT-PCR）和蛋白质印迹分析结果表明，牙龈卟啉单胞菌上清液在48小时内上调了MT1-MMP在转录和翻译水平上的表达。这些结果表明，牙龈卟啉单胞菌上清液可激活HPDL细胞中的MMP-2，其激活机制可能涉及MT1-MMP量的增加。牙龈卟啉单胞菌对MMP-2的激活可能在慢性牙周炎的过程中起作用。
