Relationships between inducing genes of nitric oxide synthase and apoptosis in the inflammatory gingival tissue.

一氧化氮合酶诱导基因与炎症牙龈组织凋亡的关系

• 批准号: 08672177
• 负责人: ENDO Hideaki
• 金额: $ 1.47万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672177/
• 关键词: Nitric Oxide Synthase; Periodonto-pathic Bacteria; HL-60; Inflammation; Apoptosis; 一酸化窒素; スーパーオキサイドディスムターゼ; カタラーゼ; 実験的歯周炎; 間接免疫蛍光抗体法; 歯肉線維芽細胞; 蛍光抗体法

Inducing Nitric Oxide Synthase in the Rat Inflammatory Gingival TissueThe distribution of 3 kinds of nitric oxide synthase ( oriented from brain, endothelial cell and macrophage ; NOS ) has been investigated in rat healthy and experimental inflamed gingiva. The samples were examined by FITC-conjugated antibody immuno-histochemically. In the part of FITC-positive area, the 3 kinds of NOS antibody were stained in 3 different width and concentration. The anti-macrophage NOS antibody area was the widest FITC-positive area. The other 2 NOS antibody staining area were observed in the localized severe inflammatory gingival area. The polymorphonuclear leukocytes (PMNs) have constitutive NOS and in the PMNs the stimulant does not induce NOS.In the experiment, 3 kinds of mouse-monoclonal NOS antibodies reacted in the inflammatory gingiva. It was suggested that the each 3 NOS antibody has a fragment matching to the reacted area of PMNs' NOS.Induction of apoptosis in HL-60 by periodonto-pathic bacteria and nitric oxidePolymorphonuclear leukocytes gather acute inflammatory gingivalarea and after the acute phase, lymphocytes gather the chronic inflamed gingiva. HL-60 (derived from acutepremyelotic leukemia) is original cell of PMNs and lymphocyte that are infection related cells. In this study, HL-60 and periodonto-pathic bacteria were incubated with and without NOR4 that has been auto-released nitric oxide. Apoptosis cells were estimated the divided shape of nucleic acid stained by fluorescent dye ( Hoechst 33342, acridine orange and ethidium bromide). The periodont-pathic bacteria were used Porphyromonas gingivalis W50, W83, ATCC 33277, Prevotella intermedia ATOC 25611, 26261. The results were as follows ; the apoptosis cells were detected all cases, differences among those bacteria were not detected. The presence of NOR4 accelerated apoptosis cells. The total average apoptosis cells were about 12% without stimulant in 3 days incubation.

在大鼠炎症牙龈组织中诱导一氧化氮合酶研究了三种一氧化氮合酶（NOS）在健康大鼠和实验性炎症牙龈组织中的分布，分别来自脑、内皮细胞和巨噬细胞。采用fitc偶联抗体免疫组织化学检测。在fitc阳性区部分，3种NOS抗体以3种不同的宽度和浓度染色。抗巨噬细胞NOS抗体区为最宽的fitc阳性区。另外2个NOS抗体染色区位于局部重度炎症龈区。多形核白细胞（PMNs）具有组成型NOS，在PMNs中，兴奋剂不诱导NOS。实验中，3种小鼠单克隆NOS抗体在炎症牙龈中发生反应。提示3 - NOS抗体各有一个与PMNs NOS反应区相匹配的片段。牙周病原菌和一氧化氮诱导HL-60细胞凋亡，多形核白细胞聚集急性炎症龈区，急性期后淋巴细胞聚集慢性炎症龈区。HL-60（来源于急性髓前白血病）是PMNs和淋巴细胞的原始细胞，是感染相关细胞。在这项研究中，将HL-60和牙周病细菌与NOR4一起培养，NOR4已自动释放一氧化氮。用荧光染料（Hoechst 33342、吖啶橙和溴化乙锭）染色核酸，估计凋亡细胞的分裂形状。牙周致病菌为牙龈卟啉单胞菌W50、W83、ATCC 33277、中间普雷沃氏菌ATOC 25611、26261。实验结果如下：所有病例均检测到凋亡细胞，细菌间无差异。NOR4的存在加速了细胞凋亡。在无兴奋剂的情况下，培养3 d细胞平均凋亡细胞总数约为12%。