Wang H Y, Liu J W, Li Q, Tan L S, Lin L, Pan Y P
Department of Periodontics, School of Stomatology, China Medical University & Liaoning Institute of Dental Research, Shenyang 110002, China.
Department of Oral Biology, School of Stomatology, China Medical University & Liaoning Institute of Dental Research, Shenyang 110002, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2018 Mar 9;53(3):150-156. doi: 10.3760/cma.j.issn.1002-0098.2018.03.002.
To detect the inhibitory ability of histatin 5 on the auto-aggregation of (Pg), and the co-aggregation of Pg with (Fn); and to provide a theoretical basis for the role of oral innate immunity played in the inhibition of chronic periodontitis. Saliva and supragingival, subgingival plaque samples were collected from 49 chronic periodontitis patients in School of Stomatology, China Medical University and 27 periodontal healthy individuals. Enzyme linked immunosorbent assay was used to assess the amount of histatin 5 in saliva, absolute quantitative real-time PCR (qPCR) was applied to detect the DNA copies of Fn, Pg and total bacteria in supragingival and subgingival plaque samples. The effects of histatin 5 on auto- and co-aggregation were assessed by bacterial adhesion test and scanning electron microscopy. Hemagglutinin gene, arginine-gingipains gene in Pg and FomA gene in Fn were tested by relative qPCR. Independent samples -test was used to calculate the significance between the experimental group and the control group. -value<0.05 was considered statistically significant. For chronic periodontitis patients, there was an inverse correlation between the concentration of histatin 5 and Fn and Pg in supragingival plaque samples (-0.379, -0.624). Similarly, an inverse correlation was also observed between the concentration of histatin 5 and subgingival Fn and Pg, respectively (-0.404, -0.314). As for periodontally healthy individuals, there was an inverse correlation between the concentration of histatin 5 and supragingival and subgingival Pg (-0.572, -0.533). Bacterial adhesion test and scanning electron microscopy certified that 25 mg/L histatin 5 inhibited the auto-aggregation of Pg-Pg and the co-aggregation of Pg-Fn. Results of qPCR showed that 25 mg/L histatin 5 up-regulated hemagglutinin gene by (14.52±3.25) fold and down-regulated FomA gene to (0.22±0.10) fold. Histatin 5 could inhibit the auto-aggregation of Pg-Pg and the co-aggregation of Pg-Fn by regulating hemagglutinin gene and FomA gene expression.
检测组蛋白5对牙龈卟啉单胞菌（Pg）自聚集以及Pg与具核梭杆菌（Fn）共聚集的抑制能力；为口腔天然免疫在抑制慢性牙周炎中所起的作用提供理论依据。收集中国医科大学口腔医学院49例慢性牙周炎患者及27例牙周健康个体的唾液、龈上和龈下菌斑样本。采用酶联免疫吸附测定法评估唾液中组蛋白5的含量，应用绝对定量实时聚合酶链反应（qPCR）检测龈上和龈下菌斑样本中Fn、Pg及总细菌的DNA拷贝数。通过细菌黏附试验和扫描电子显微镜评估组蛋白5对自聚集和共聚集的影响。采用相对qPCR检测Pg中的血凝素基因、精氨酸牙龈蛋白酶基因以及Fn中的FomA基因。使用独立样本t检验计算实验组与对照组之间的显著性差异。P值<0.05被认为具有统计学意义。对于慢性牙周炎患者，龈上菌斑样本中组蛋白5浓度与Fn和Pg之间呈负相关（-0.379，-0.624）。同样，组蛋白5浓度与龈下Fn和Pg之间也分别呈负相关（-0.404，-0.314）。对于牙周健康个体，组蛋白5浓度与龈上和龈下Pg之间呈负相关（-0.572，-0.533）。细菌黏附试验和扫描电子显微镜证实，25 mg/L组蛋白5可抑制Pg - Pg的自聚集以及Pg - Fn的共聚集。qPCR结果显示，25 mg/L组蛋白5使血凝素基因上调（14.52±3.25）倍，使FomA基因下调至（0.22±0.10）倍。组蛋白5可通过调节血凝素基因和FomA基因表达来抑制Pg - Pg的自聚集以及Pg - Fn的共聚集。
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