Induction of murine interleukin-1 beta expression by water-soluble components from Hericium erinaceum.
OBJECTIVE
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.


CONCLUSIONS
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.
Son
Chang-Gue
CG
East-West Cancer Center, Dunsan Oriental Hospital of Daejon University, Daejon, 301-724, Republic of Korea.
Shin
Jang-Woo
JW
Cho
Jung-Hyo
JH
Cho
Chong-Kwan
CK
Yun
Cheol-Heui
CH
Han
Seung-Hyun
SH
D000363:Agaricales / Q000737:chemistry; D000818:Animals; D001688:Biological Products / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D053583:Interleukin-1beta / Q000096:biosynthesis* / Q000235:genetics; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008809:Mice, Inbred C3H; D016328:NF-kappa B / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D018808:Transcription Factor AP-1 / Q000378:metabolism; D047071:beta-Glucans / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*
D001688:Biological Products; D022782:CCAAT-Enhancer-Binding Protein-beta; D053583:Interleukin-1beta; D016328:NF-kappa B; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; D047071:beta-Glucans