Oral & Dental Sciences Research Group, School of Postgraduate Medical and Dental Education, University of Central Lancashire, Preston, UK.
J Alzheimers Dis. 2013;36(4):665-77. doi: 10.3233/JAD-121918.
The aim of this study was to establish a link between periodontal disease and Alzheimer's disease (AD) with a view to identifying the major periodontal disease bacteria (Treponema denticola, Tannerella forsythia, and Porphyromonas gingivalis) and/or bacterial components in brain tissue from 12 h postmortem delay. Our request matched 10 AD cases for tissue from Brains for Dementia Research alongside 10 non-AD age-related controls with similar or greater postmortem interval. We exposed SVGp12, an astrocyte cell line, to culture supernatant containing lipopolysaccharide (LPS) from the putative periodontal bacteria P. gingivalis. The challenged SVGp12 cells and cryosections from AD and control brains were immunolabeled and immunoblotted using a battery of antibodies including the anti-P. gingivalis-specific monoclonal antibody. Immunofluorescence labeling demonstrated the SVGp12 cell line was able to adsorb LPS from culture supernatant on its surface membrane; similar labeling was observed in four out of 10 AD cases. Immunoblotting demonstrated bands corresponding to LPS from P. gingivalis in the SVGp12 cell lysate and in the same four AD brain specimens which were positive when screened by immunofluorescence. All controls remained negative throughout while the same four cases were consistently positive for P. gingivalis LPS (p = 0.029). This study confirms that LPS from periodontal bacteria can access the AD brain during life as labeling in the corresponding controls, with equivalent/longer postmortem interval, was absent. Demonstration of a known chronic oral-pathogen-related virulence factor reaching the human brains suggests an inflammatory role in the existing AD pathology.
本研究旨在建立牙周病与阿尔茨海默病(AD)之间的联系,以期确定主要的牙周病细菌(齿密螺旋体、福赛斯坦纳菌和牙龈卟啉单胞菌)和/或脑组织中 12 小时死后延迟的细菌成分。我们的要求是为痴呆症研究大脑组织匹配 10 例 AD 病例,以及 10 例非 AD 年龄相关性对照,其死后间隔时间相似或更长。我们使 SVGp12 (星形胶质细胞系)暴露于来自假定牙周细菌牙龈卟啉单胞菌的培养上清液中的脂多糖(LPS)中。受挑战的 SVGp12 细胞和 AD 及对照大脑的冷冻切片用一系列抗体进行免疫标记和免疫印迹,包括抗牙龈卟啉单胞菌特异性单克隆抗体。免疫荧光标记表明 SVGp12 细胞系能够在其细胞膜表面吸附来自培养上清液的 LPS;在 10 例 AD 病例中的 4 例中观察到类似的标记。免疫印迹显示在 SVGp12 细胞裂解物和在免疫荧光筛选为阳性的相同的 4 个 AD 脑标本中存在与牙龈卟啉单胞菌 LPS 对应的条带。所有对照在整个过程中均为阴性,而相同的 4 个病例始终对牙龈卟啉单胞菌 LPS 呈阳性(p = 0.029)。本研究证实,牙周病细菌的 LPS 可以在生命过程中进入 AD 大脑,因为在具有相同死后间隔的相应对照中不存在标记。已知慢性口腔病原体相关毒力因子到达人脑的证明表明其在现有 AD 病理学中具有炎症作用。