Induction of matrix metalloproteinase-9 in alveolar macrophages by TNF-α through NF-κB signal pathwa

来源 :Journal of Nanjing Medical University | 被引量 : 0次 | 上传用户:pjkxqx
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Objective: To explore the effect of tumor necrosis factor (TNF)-α on matrix metalloproteinase-9 (MMP-9) expression and activity in alveolar macrophages (AM) from patients with chronic obstructive pulmonary disease (COPD) and study its associated signal pathway. Methods: AM were collected from bronchoalveolar lavage fluid in patients with COPD. The AM were incubated for 1.5 h with pyrrolidine dithiocarbamate(PDTC)at concentrations from 0 μmol/L to 50 μmol/L and then stimulated for 24 h by TNF-α at 10 ng/ml. MMP-9 expression and activity were respectively detected by semi- quantitative reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and Zymography. NF-κB activity was investigated by electrophoretic mobility shift assay (EMSA). Results: Both the mRNA and protein levels of MMP-9 induced by TNF-α in AM were significantly elevated in a dose dependent manner (P<0.05). The level of MMP-9 activity was also correspondingly significantly elevated in the induction (P<0.05), which was possibly related with the over- expression of MMP-9. NF-κB activity was significantly increased when AM were stimulated by 10 ng/mL TNF-α (P<0.05). The expression of MMP-9 induced by TNF-α could be significantly inhibited by PDTC (P< 0.05). Conclusion: The expression and activity of MMP-9 from AM could be induced by TNF-α, and NF-κB signal pathway played an important role in the induction. Objective: To explore the effect of tumor necrosis factor (TNF) -α on matrix metalloproteinase-9 (MMP-9) expression and activity in alveolar macrophages (AM) from patients with chronic obstructive pulmonary disease (COPD) and study its associated signal pathway . AM: AM were collected from bronchoalveolar lavage fluid in patients with COPD. The AM were incubated for 1.5 h with pyrrolidine dithiocarbamate (PDTC) at concentrations from 0 μmol / L to 50 μmol / L and then stimulated for 24 h by TNF-α at 10 ng / ml. MMP-9 expression and activity were detected respectively by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and Zymography. NF-κB activity was investigated by electrophoretic mobility shift assay (EMSA) Results: Both the mRNA and protein levels of MMP-9 induced by TNF-α in AM were significantly elevated in a dose dependent manner (P <0.05). The level of MMP-9 activity was also correspondingly significantly elevated in the induction (P <0.05), which was possibly related to the over-expression of MMP-9. The expression of MMP-9 was significantly increased when AM was stimulated by 10 ng / mL TNF- 9 induced by TNF-α could be inhibited by PDTC (P <0.05). Conclusion: The expression and activity of MMP-9 from AM could be induced by TNF-α, and NF-κB signal pathway played an important role in the induction.
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