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Induction of murine interleukin-1 beta expression by water-soluble components from Hericium erinaceum
Chang-Gue Son1, Jang-Woo Shin, Jung-Hyo Cho
1East-West Cancer Center, Dunsan Oriental Hospital of Daejon University, Daejon, 301-724, Republic of Korea.
Insights
Water extract from Hericium erinaceum (WEHE) boosts interleukin-1beta (IL-1beta) expression in macrophages. This occurs at the gene transcription level by activating key transcription factors, including NF-kappaB.
Area of Science:
- Immunology
- Molecular Biology
- Pharmacology
Background:
- Interleukin-1beta (IL-1beta) is a key inflammatory cytokine.
- Hericium erinaceum (WEHE) is being investigated for its potential biological activities.
Purpose of the Study:
- To investigate the inductive effect of water extract from Hericium erinaceum (WEHE) on interleukin-1beta (IL-1beta) expression in macrophages.
Main Methods:
- Murine macrophage cell-line (RAW 264.7) stimulation with WEHE.
- Analysis of IL-1beta mRNA and protein expression via RT-PCR, Western blotting, and bioassays.
- Investigation of gene regulation using CAT reporter gene assays and electrophoretic mobility shift assays (EMSA) for transcription factors (NF-kappaB, AP-1, NF-IL6, CRE/ATF).
Main Results:
- WEHE dose-dependently induced IL-1beta mRNA and protein production.
- WEHE enhanced IL-1beta gene transcription.
- WEHE increased the binding activity of NF-kappaB, AP-1, and NF-IL6 transcription factors.
Conclusions:
- WEHE induces IL-1beta expression in macrophages.
- The induction occurs at the transcriptional level.
- WEHE enhances IL-1beta expression by activating NF-kappaB, NF-IL6, and AP-1 transcription factors.
Aim:
To investigate the inductive effect of water extract from Hericium erinaceum (WEHE) on interleukin-1beta (IL-1beta) expression.
Methods:
A murine macrophage cell-line, RAW 264.7 was stimulated with 1 to 10 mg/L WEHE and inductions of IL-1beta 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-1beta 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-1beta promoter. An electrophoretic mobility shift assay (EMSA) was also performed to examine transcription factors, nuclear factor-kappa B (NF-kappaB), activator protein 1 (AP-1), nuclear factor interleukin-6 (NF-IL6), and cAMP response element (CRE)/activating transcription factor (ATF).
Results:
WEHE induced IL-1beta production in both its mRNA and protein expression in a dose-dependent manner. The inductive effect of WEHE on IL-1beta gene expression was due to the augmentation of the IL-1beta transcription. Furthermore, EMSA showed that WEHE markedly increased the binding activities of NF-kappaB, 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-1beta gene expression.
Conclusion:
WEHE induces IL-1kappa expression in macrophages at a transcriptional level by enhancing the activation of transcription factors, NF-kappaB, NF-IL6, and AP-1.
