Induction of murine interleukin-1 beta expression by water-soluble components from Hericium erinaceum
Abstract
Aim: To investigate the inductive effect of water extract from Hericium erinaceum (WEHE) on interleukin-1 β (IL-1β) expression.
Methods: A murine macrophage cell-line, RAW 264.7 was stimulated with 1 to 10 mg/L WEHE and inductions of IL-1β protein and its steady state mRNA were examined using a bioassay, Western blotting, and reverse transcription-polymerase chain reaction (RT-PCR) analysis. The inductive effect of WEHE on IL-1β gene expression was further investigated by a chloramphenicol acetyltransferase (CAT) reporter gene assay using a transient transfection with pIL-1(870 bp)-CAT where the expression of the CAT gene was regulated by a IL-1β promoter. An electrophoretic mobility shift assay (EMSA) was also performed to examine transcription factors, nuclear factor-κ B (NF-κB), activator protein 1 (AP-1), nuclear factor interleukin-6 (NF-IL6), and cAMP response element (CRE)/activating transcription factor (ATF).
Results: WEHE induced IL-1β production in both its mRNA and protein expression in a dose-dependent manner. The inductive effect of WEHE on IL-1β gene expression was due to the augmentation of the IL-1β transcription. Furthermore, EMSA showed that WEHE markedly increased the binding activities of NF-κB, and to a lesser extent, those of AP-1 and NF-IL6 to their cognate DNA recognition sites, whereas CRE/ATF binding remained constant, all of which are known to be involved in the regulation of IL-1β gene expression.
Conclusion: WEHE induces IL-1β expression in macrophages at a transcriptional level by enhancing the activation of transcription factors, NF-κB, NF-IL6, and AP-1.
Keywords:
Methods: A murine macrophage cell-line, RAW 264.7 was stimulated with 1 to 10 mg/L WEHE and inductions of IL-1β protein and its steady state mRNA were examined using a bioassay, Western blotting, and reverse transcription-polymerase chain reaction (RT-PCR) analysis. The inductive effect of WEHE on IL-1β gene expression was further investigated by a chloramphenicol acetyltransferase (CAT) reporter gene assay using a transient transfection with pIL-1(870 bp)-CAT where the expression of the CAT gene was regulated by a IL-1β promoter. An electrophoretic mobility shift assay (EMSA) was also performed to examine transcription factors, nuclear factor-κ B (NF-κB), activator protein 1 (AP-1), nuclear factor interleukin-6 (NF-IL6), and cAMP response element (CRE)/activating transcription factor (ATF).
Results: WEHE induced IL-1β production in both its mRNA and protein expression in a dose-dependent manner. The inductive effect of WEHE on IL-1β gene expression was due to the augmentation of the IL-1β transcription. Furthermore, EMSA showed that WEHE markedly increased the binding activities of NF-κB, and to a lesser extent, those of AP-1 and NF-IL6 to their cognate DNA recognition sites, whereas CRE/ATF binding remained constant, all of which are known to be involved in the regulation of IL-1β gene expression.
Conclusion: WEHE induces IL-1β expression in macrophages at a transcriptional level by enhancing the activation of transcription factors, NF-κB, NF-IL6, and AP-1.