Abstract
In order to explore the role of Krupperl-like factor 4(KLF4) in regulating the expression of IL-6 induced by lipopolysaccharide(LPS).mRNA and protein levels of KLF4 in various periods were determined by RT-PCR and Western blot.A full-length cDNA or antisense oligonucleotides of KLF4 was transfected into RAW264.7 macrophages.The expression and release of IL-6 were analyzed by RT-PCR and ELISA.The transcription and DNA binding activities of KLF4 to the IL-6 promoter were detected by the luciferase reporter and EMSA.The results showed that treatment of RAW264.7 macrophages with LPS resulted in increase in KLF4 and IL-6 protein levels;KLF4 overexpression decreased the expression and release of IL-6, while KLF4 inhibition increased the expression of IL-6.The results from luciferase reporter gene assay demonstrated that KLF4 could inhibit the transcriptional activity of IL-6 gene.The results from EMSA demonstrated that KLF4 could not bind to the KLF4 binding sites on the promoter of IL-6 gene.These data suggested that LPS can stimulated the expression of KLF4 and release of IL-6 in RAW264.7 macrophages;KLF4 can inhibited the release of IL-6 in LPS-stimulated RAW264.7 macrophages;KLF4 can inhibited the transcription activity of IL-6 promoter in RAW264.7 macrophages;this effect of KLF4 is not through binding the IL-6 promoter directly.
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