节点文献
吡咯烷二硫代氨基甲酸盐下调基质金属蛋白酶9表达机制的研究
Study on the mechanism of pyrrolidine dithiocarbamate down-regulating matrix metalloproteinase-9 expression
【摘要】 目的探讨TNF-α诱导的肺泡巨噬细胞(AM)性基质金属蛋白酶9(MMP-9)表达的信号通路以及吡咯烷二硫代氨基甲酸盐(PDTC)对MMP-9表达的影响机制。方法从慢性阻塞性肺疾病患者支气管肺泡灌洗液中分离与培养AM,以PDTC预处理AM,以TNF-α或IL-1刺激AM。半定量逆转录-聚合酶链反应法检测MMP-9 mRNA的表达;Western blot检测MMP-9蛋白的表达及TNF-α或IL-1诱导的IκBα磷酸化水平。凝胶阻滞分析实验检测NF-κB活性。结果TNF-α上调AM源性MMP-9 mRNA和蛋白的表达(P<0.05);PDTC抑制TNF-α诱导的MMP-9的表达(P<0.05)。PDTC对TNF-α或IL-1诱导的IκBα的磷酸化均无抑制作用(P>0.05)。PDTC对TNF-α或IL-1诱导的NF-κB的活化均有抑制作用(P<0.05);且PDTC不能在体外直接抑制NF-κB的DNA结合活性(P>0.05)。结论NF-κB在TNF-α诱导的AM源性MMP-9的表达中起着重要作用;PDTC可能通过抑制泛素化-蛋白酶小体途径来下调TNF-α诱导的AM源性MMP-9的表达。
【Abstract】 Aim To study the effects of pyrrolidine dithiocarbamate(PDTC) on matrix metalloproteinase-9(MMP-9) expression in alveolar macrophages(AM) induced by TNF-α,and then to explore it’s signal pathway.Methods AM were collected from bronchoalveolar lavage fluid in patients with chronic obstructive pulmonary disease.AM were pretreated with PDTC and then stimulated by TNF-α or IL-1.MMP-9 mRNA expression was detected by semi-quantitative reverse transcription-polymerase chain reaction and MMP-9 protein expression was detected by Western blot.Phospho-IκBα protein levels induced by TNF-α or IL-1 were detected by Western blot.NF-κB activity was detected by electrophoretic mobility shift assay.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 expression of MMP-9 induced by TNF-α was significantly inhibited by PDTC(P<0.05).PDTC had no clear effect of inhibiting the phosphorylation of IκBα induced by TNF-α or IL-1(P>0.05).PDTC had distinct effect of inhibiting the activation of NF-κB induced by TNF-α or IL-1 in a dose-dependent manner(P<0.05).It also showed PDTC couldn’t directly inhibit NF-κB DNA binding activity in vitro(P>0.05).Conclusions NF-κB plays an important role in MMP-9 expression induced by TNF-α in AM;PDTC can down-regulate MMP-9 expression induced by TNFα in AM possibly through preventing the degradation of IκBα via ubiquitylation-proteasome proteolytic pathway.
【Key words】 nuclear factor-kappaB; pyrrolidine dithiocarbamate; matrix metalloproteinase; IκBα;
- 【文献出处】 中国药理学通报 ,Chinese Pharmacological Bulletin , 编辑部邮箱 ,2009年01期
- 【分类号】R96
- 【被引频次】11
- 【下载频次】92