Haraszthy V I, Zambon J J, Trevisan M, Zeid M, Genco R J
Department of Oral Biology, School of Dental Medicine, State University of New York at Buffalo, USA.
J Periodontol. 2000 Oct;71(10):1554-60. doi: 10.1902/jop.2000.71.10.1554.
Recent studies suggest that chronic infections including those associated with periodontitis increase the risk for coronary vascular disease (CVD) and stroke. We hypothesize that oral microorganisms including periodontal bacterial pathogens enter the blood stream during transient bacteremias where they may play a role in the development and progression of atherosclerosis leading to CVD.
To test this hypothesis, 50 human specimens obtained during carotid endarterectomy were examined for the presence of Chlamydia pneumoniae, human cytomegalovirus, and bacterial 16S ribosomal RNA using specific oligonucleotide primers in polymerase chain reaction (PCR) assays. Approximately 100 ng of chromosomal DNA was extracted from each specimen and then amplified using standard conditions (30 cycles of 30 seconds at 95 degrees C, 30 seconds at 55 degrees C, and 30 seconds at 72 degrees C). Bacterial 16S rDNA was amplified using 2 synthetic oligonucleotide primers specific for eubacteria. The PCR product generated with the eubacterial primers was transferred to a charged nylon membrane and probed with digoxigenin-labeled synthetic oligonucleotides specific for Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, and Prevotella intermedia.
Eighty percent of the 50 endarterectomy specimens were positive in 1 or more of the PCR assays. Thirty-eight percent were positive for HCMV and 18% percent were positive for C. pneumoniae. PCR assays for bacterial 16S rDNA also indicated the presence of bacteria in 72% of the surgical specimens. Subsequent hybridization of the bacterial 16S rDNA positive specimens with species-specific oligonucleotide probes revealed that 44% of the 50 atheromas were positive for at least one of the target periodontal pathogens. Thirty percent of the surgical specimens were positive for B. forsythus, 26% were positive for P. gingivalis, 18% were positive for A. actinomycetemcomitans, and 14% were positive for P. intermedia. In the surgical specimens positive for periodontal pathogens, more than 1 species was most often detected. Thirteen (59%) of the 22 periodontal pathogen-positive surgical specimens were positive for 2 or more of the target species.
Periodontal pathogens are present in atherosclerotic plaques where, like other infectious microorganisms such as C. pneumoniae, they may play a role in the development and progression of atherosclerosis leading to coronary vascular disease and other clinical sequelae.
近期研究表明，包括与牙周炎相关的慢性感染会增加患冠状动脉血管疾病（CVD）和中风的风险。我们推测，包括牙周细菌病原体在内的口腔微生物会在短暂菌血症期间进入血流，在那里它们可能在导致CVD的动脉粥样硬化的发生和发展中发挥作用。
为验证这一推测，在颈动脉内膜切除术期间获取的50份人体标本，使用聚合酶链反应（PCR）检测中的特异性寡核苷酸引物，检测肺炎衣原体、人巨细胞病毒和细菌16S核糖体RNA的存在情况。从每份标本中提取约100 ng的染色体DNA，然后使用标准条件（95℃30秒、55℃30秒、72℃30秒，共30个循环）进行扩增。使用2种针对真细菌的合成寡核苷酸引物扩增细菌16S rDNA。用真细菌引物产生的PCR产物转移到带电荷的尼龙膜上，并用针对伴放线放线杆菌、福赛坦氏菌、牙龈卟啉单胞菌和中间普氏菌的地高辛标记合成寡核苷酸进行探针杂交。
50份内膜切除术标本中有80%在1种或更多种PCR检测中呈阳性。38%的标本HCMV呈阳性，18%的标本肺炎衣原体呈阳性。细菌16S rDNA的PCR检测也表明72%的手术标本中有细菌存在。随后，将细菌16S rDNA阳性标本与种特异性寡核苷酸探针杂交，结果显示50个动脉粥样硬化斑块中有44%至少对一种目标牙周病原体呈阳性。30%的手术标本福赛坦氏菌呈阳性，26%的标本牙龈卟啉单胞菌呈阳性，18%的标本伴放线放线杆菌呈阳性，14%的标本中间普氏菌呈阳性。在牙周病原体呈阳性的手术标本中，最常检测到不止1种病原体。22份牙周病原体阳性手术标本中有13份（59%）对2种或更多种目标菌种呈阳性。
牙周病原体存在于动脉粥样硬化斑块中，与肺炎衣原体等其他感染性微生物一样，它们可能在导致冠状动脉血管疾病和其他临床后遗症的动脉粥样硬化的发生和发展中发挥作用。
