Kuula Heidi, Salo Tuula, Pirilä Emma, Hagström Jaana, Luomanen Marita, Gutierrez-Fernandez Ana, Romanos Georgios E, Sorsa Timo
Biomedicum Helsinki, Institute of Dentistry, University of Helsinki, PO Box 63, FIN-00014 Helsinki, Finland.
Arch Oral Biol. 2008 Feb;53(2):175-86. doi: 10.1016/j.archoralbio.2007.09.010. Epub 2007 Nov 9.
Aberrant matrix metalloproteinase (MMP) and human beta-defensin (HBD) functions have been found in inflammatory diseases. The objectives of this study were to investigate the immunolocalisation, mRNA expression and molecular forms of MMP-25, MMP-26, HBD-1 and HBD-2 in chronic and aggressive periodontitis and in peri-implantitis. The expression of MMP-25 by cultured human plasmacytoma cells and macrophages, and the effects of MMP-26 and Porphyromonas gingivalis trypsin-like proteinase on HBD-1 and -2 were also studied.
Immunohistochemistry, immunofluorescent analysis, reverse transcriptase polymerase chain reaction and immunoblotting were used to assess localisation, mRNA expression and molecular forms of MMP-25, MMP-26, HBD-1 and HBD-2. HBD degradation by MMP-26 and P. gingivalis proteinase was studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.
MMP-25 was present in plasma cells and polymorphonuclear leucocytes, and MMP-26 was present in oral and sulcular basement membrane zones. HBD-1 was distributed perivasculary in gingival connective tissue and in oral and sulcular epithelium, and HBD-2 was found to a lesser extent in the perivascular space. Low MMP-25, MMP-26, HBD-1 and HBD-2 mRNA expression was found. Immunoblot revealed 29-57-kDa MMP-25 in myeloma cell lysates, but not in macrophages, and partly activated MMP-25 and -26 in diseased gingival crevicular fluid and peri-implant sulcular fluid. P. gingivalis trypsin-like proteinase degraded HBD-1 and -2.
Both MMP-25 and -26 were expressed more strongly in extensively inflamed gingiva compared with healthy gingiva. The expression of HBD-1 was stronger than that of HBD-2 in periodontitis and peri-implantitis. De-novo expression of MMP-25 and -26 is associated with periodontal and peri-implant inflammation. Furthermore, P. gingivalis trypsin-like proteinase, but not MMP-26, can degrade HBD-1 and -2, which could lead to a weakened innate immune response.
在炎症性疾病中已发现基质金属蛋白酶（MMP）和人β-防御素（HBD）功能异常。本研究的目的是调查MMP-25、MMP-26、HBD-1和HBD-2在慢性和侵袭性牙周炎以及种植体周围炎中的免疫定位、mRNA表达和分子形式。还研究了培养的人浆细胞瘤细胞和巨噬细胞中MMP-25的表达，以及MMP-26和牙龈卟啉单胞菌胰蛋白酶样蛋白酶对HBD-1和-2的影响。
采用免疫组织化学、免疫荧光分析、逆转录聚合酶链反应和免疫印迹法评估MMP-25、MMP-26、HBD-1和HBD-2的定位、mRNA表达和分子形式。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳研究MMP-26和牙龈卟啉单胞菌蛋白酶对HBD的降解作用。
MMP-25存在于浆细胞和多形核白细胞中，MMP-26存在于口腔和龈沟基底膜区。HBD-1在牙龈结缔组织的血管周围以及口腔和龈沟上皮中分布，而HBD-2在血管周围间隙中的含量较少。发现MMP-25、MMP-26、HBD-1和HBD-2的mRNA表达较低。免疫印迹显示骨髓瘤细胞裂解物中有29 - 57 kDa的MMP-25，但巨噬细胞中没有，并且在患病的龈沟液和种植体周围龈沟液中有部分活化的MMP-25和-26。牙龈卟啉单胞菌胰蛋白酶样蛋白酶可降解HBD-1和-2。
与健康牙龈相比，MMP-25和-26在广泛炎症的牙龈中表达更强。在牙周炎和种植体周围炎中，HBD-1的表达强于HBD-2。MMP-25和-26的从头表达与牙周和种植体周围炎症相关。此外，牙龈卟啉单胞菌胰蛋白酶样蛋白酶而非MMP-26可降解HBD-1和-2，这可能导致固有免疫反应减弱。
