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

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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.