Sasaki Haruka, Watanabe Kiyoko, Toyama Toshizo, Koyata Yasunori, Hamada Nobushiro
Department of Microbiology, Kanagawa Dental University, Yokosuka, Kanagawa 238-8580, Japan.
J Vet Med Sci. 2015 Mar;77(3):265-71. doi: 10.1292/jvms.14-0463. Epub 2014 Nov 25.
Porphyromonas gulae is considered to be associated with canine periodontitis. We have previously reported that the P. gulae American Type Culture Collection (ATCC) 51700 comprised 41-kDa fimbriae. The purpose of the present study was to demonstrate the roles of 41-kDa fimbrial protein in periodontal disease. In this study, we examined the involvement of the 41-kDa fimbrial protein in osteoclast differentiation and cytokine production in murine macrophages. Furthermore, alveolar bone resorption induced by P. gulae infection in rats was evaluated. To estimate osteoclast differentiation, bone marrow cells and MC3T3-G2/PA6 cells were cultured with or without the 41-kDa fimbrial protein for 7 days. BALB/c mouse peritoneal macrophages were stimulated with the 41-kDa fimbrial protein, and the levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α production were determined by enzyme-linked immunosorbent assay. Osteoclast differentiation was significantly enhanced by treatment with the 41-kDa fimbrial protein in a dose-dependent manner. The total area of pits formed on the dentine slices with osteoclasts incubated with the 41-kDa fimbrial protein was significantly greater than that of the control. The purified 41-kDa fimbrial protein induced IL-1β and TNF-α production in BALB/c mouse peritoneal macrophages after 6 hr of incubation in a dose-dependent manner. The bone loss level in rats infected with P. gulae was significantly higher than that of the sham-infected rats. These results suggest that P. gulae 41-kDa fimbriae play important roles in the pathogenesis of periodontal disease.
牙龈卟啉单胞菌被认为与犬牙周炎有关。我们之前报道过牙龈卟啉单胞菌美国模式培养物集存库(ATCC)51700含有41-kDa菌毛。本研究的目的是证明41-kDa菌毛蛋白在牙周疾病中的作用。在本研究中,我们检测了41-kDa菌毛蛋白在小鼠巨噬细胞破骨细胞分化和细胞因子产生中的作用。此外,还评估了牙龈卟啉单胞菌感染诱导的大鼠牙槽骨吸收情况。为了评估破骨细胞分化,将骨髓细胞和MC3T3-G2/PA6细胞在有或无41-kDa菌毛蛋白的情况下培养7天。用41-kDa菌毛蛋白刺激BALB/c小鼠腹腔巨噬细胞,通过酶联免疫吸附测定法测定白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的产生水平。用41-kDa菌毛蛋白处理可显著增强破骨细胞分化,且呈剂量依赖性。与41-kDa菌毛蛋白一起孵育的破骨细胞在牙本质切片上形成的凹坑总面积显著大于对照组。纯化的41-kDa菌毛蛋白在孵育6小时后以剂量依赖性方式诱导BALB/c小鼠腹腔巨噬细胞产生IL-1β和TNF-α。感染牙龈卟啉单胞菌的大鼠的骨丢失水平显著高于假感染大鼠。这些结果表明牙龈卟啉单胞菌41-kDa菌毛在牙周疾病的发病机制中起重要作用。