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

本实验观察了大鼠正常牙龈和实验性牙龈炎组织中一氧化氮合酶（NOS）的分布。用FITC标记的抗体进行免疫组化检测。在FITC阳性区域，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2个NOS抗体染色区位于局部重度炎症牙龈区。多形核白细胞（PMNs）具有组成型NOS，刺激剂对PMNs中的NOS无诱导作用。牙周致病菌和一氧化氮对HL-60细胞凋亡的诱导作用：多形核白细胞聚集于急性炎症区，急性期后淋巴细胞聚集于慢性炎症区。HL-60（来源于急性早幼粒细胞白血病）是感染相关的中性粒细胞和淋巴细胞的原始细胞。在这项研究中，HL-60和牙周病细菌孵育与NOR 4，已自动释放一氧化氮。用Hoechst 33342、吖啶橙子和溴化乙锭荧光染色，观察细胞凋亡的核酸分裂形态。牙周病致病菌是牙龈卟啉单胞菌W50、W83、ATCC 33277、中间普雷沃氏菌ATOC 25611、26261。结果表明：所有病例均检测到凋亡细胞，各细菌间无差异。NOR 4的存在促进细胞凋亡。在无刺激的情况下，培养3天，凋亡细胞总数平均约为12%。

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.