诱导型一氧化氮合酶_

核因子κB对人单核细胞诱导型一氧化氮合酶表达的调控作用

http://www.100md.com 《第四军医大学学报》 2004年第12期

诱导型一氧化氮合酶,,核因子κB；诱导型一氧化氮合酶；基因表达,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Nuclear factor κB regulates gene expression of inducible nitricoxide synthase in human monocyte

    ZHANG Ning, XU YongJian, ZHANG ZhenXiang

    Department of Respiratory Medicine, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430030, China

    【Abstract】 AIM: To investigate whether gene expression of inducible nitric oxide synthase(iNOS)in human monocyte is modulated by nuclear factor κB(NFκB). METHODS: Human monocytes were cultured in medium with lipopolysaccharide(LPS)± pyrrolidine dithiocarbamate(PDTC)or in medium alone. One hour later, the nuclear protein of the monocytes was extracted for electrophoretic mobility shift assay (EMSA) and the monocyte coverslips were collected for immunofluorescence staining for NFκB p65 subunit. Ten hours later, RNA was extracted for RTPCR for iNOS. Twentyfour hours later, whole cell extracts were used for Western blot and monocyte corverslips were collected for immunocytochemical staining. RESULTS: LPS increased the NFκB activation compared with that of the unstimulated cells. Ten hours later, the mRNA expression of iNOS increased and twentyfour hours later the protein expression of iNOS increased markedly. The inhibition of NFκB activation by PDTC resulted in a significant decrease of the expressions of iNOS mRNA and protein hours later. The positive rate of nuclear staining of p65 and the DNA binding activity of NFκB significantly were correlated with the mRNA and protein expression of iNOS. CONCLUSION: NFκB regulates the gene expression of iNOS in human monocytes. ......

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