| Introduction: A majority of pulmonary diseases, such as lung hurt, asthma, chronic obstructive pulmonary disease (COPD), have strong affinity with immune and inflammatory responses. Nuclear factor-KB (NF-?B) is an important pluripotent transcription factor. It's gene products include cytokine, chemokines, cell adhesion molecules, and immunoreceptors. NF-?B plays a key role in immune and inflammatory responses. Therefore, modulation of NF-KB activation may provide a direct way of inhibiting inflammatory mediators. Directing drug discovery efforts towards NF-KB activation rather than towards any one of its many target genes could produce a much greater therapeutic benefit by inhibiting expression of the constellation of NF-?B-induced pro-inflammatory genes.Eucalyptus globulus oil shows particular anti-inflammation properties, which include two main components: 1.8-cineole and a-pinene. However, the molecular of the anti-inflammation activity of eucalyptus globulus oil is still not well understood. Since NF-KB is a central mediator of inflammatory responses, we hypothesize that NF-KB is a target for eucalyptus globulus oil-mediated anti-inflammation properties.Objective: Set up Lipopolysaccharide (LPS)-induced THP-1 cells inflammation model and rats' chronic lung inflammation models induced respectively by LPS-airway-injection and sulfur dioxide (S02)-airway-inhalation. Detect the activation of nuclear factor-?B. Discuss the effects of Eucalyptus globulus oil and its main component, a-pinene on the activation of nuclear factor-KB of the three above models.Methods: THP-1 cells were incubated with a series of doses of drugs before being stimulated with LPS (1 mg.L-1, 30 min). SD rats were injected with LPS or inhaled with SO2 from airway and took a series of doses of drugs orally. The location of NF-?B p65 subunit (NF-?B/p65) in THP-1 cells and pulmonary alveolar macrophages (PAMs) was detected by indirect immunofluorescence and laser scanning confocal microscope(LSCM). The expression of NF-?B/p65 in nuclei and I?Ba in cytoplasm were measured by Western -blot analysis or Sandwich ELISA analysis. Semi-quantitative analysis of Western -blot was measured. The data are presented as mean ?SD and compared with one-way ANOVA analysis using SPSS statistical program.Rusluts:1. Indirect immunofluorescence under LSCM showed: ?The majority of FITC-label NF-?B/p65 located in the cytoplasm of control THP-1 cells or PAMs, whereas, being stimulated with LPS the majority of NF-?B/p65 located in nuclei. ?No such fluorescence were seen in the nuclei of the cells pretreated with eucalyptus globulus oil or a-pinene.2. Western-blot analysis showed: ?A low level expression of NF-?B/p65 in nuclei was measured by in control THP-1 cells or rat lung tissue, but a obvious induction of NF-?B/p65 nuclear translocation was observed in THP-1 cells being stimulated with LPS (P<0..05) or in rat lung tissue being stimulated with LPS, SO2 from airway. (2) Eucalyptus globulus oil and a-pinene (1?10?100 礸.L-1, 30 min) pretreatment decreased the NF-?B/p65 nuclear translocation in LPS-stimulated THP-1 cells, and these effects were dose-dependent (P<0.05). Eucalyptus globulus oil decreased the NF-?B/p65 nuclear translocation in the rat lung tissue being .stimulated with LPS, SOifrom airway. (? Only treatment with a-pinene (1?10?100 ng.L-1, 30 min) did not significantly affect on the NF-?B/p65 nuclear translocation in normal THP-1 cells (P>0.05). ?The I?Ba protein level in LPS-stimulated THP-1 cells markedly decreased at 30 min. The LPS-induced degradation of I?Ba was blocked by a-pinene (P<0.05).3. Sandwich ELISA analysis showed: A45onm of NF-?B/p65 in the rat lung tissue being inhaleded with SO2 from airway was markedly increased compared with control, but this effect was decreased by treated with eucalyptus globulus oil (P<0.05).Conclusions:1. In LPS-induced THP-1 cells inflammation model, LPS could stimulate the NF-icB activation. In the rats' chronic lung inflammation models induced respe... |
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