Pro-inflammatory cytokine gene expression and nitric oxide regulation of aqueous extracted Astragali radix in RAW 264.7 macrophage cells.
Astragali radix, which has tonifying and circulatory effect as well as immune response, is one of the oldest and most frequently used crude drug for oriental medicine in many Asian countries. The present study was conducted to evaluate the effect of aqueous extract of Astragali radix (ARE) on the functions of murine macrophage cell line, RAW 264.7 macrophage cells. In the cell proliferation assay, methotrexate (MTX), an agent of immune suppression, decreased the cell proliferation of RAW 264.7 macrophage cells (IC(50): 100 microg/ml), but the suppression of cell proliferation was significantly protected by ARE treatment in RAW 264.7 macrophage cells. The expressions of cytokine gene by ARE were investigated using reverse transcription polymerase chain reaction (RT-PCR). In RT-PCR, IL-1alpha, IL-1beta and IL-6 mRNA expressions was induced in ARE-treated RAW 264.7 macrophage cells. We also investigated the effect of the nitric oxide (NO) synthesis and inducible nitric oxide synthase (iNOS) mRNA expression by ARE. ARE alone had no effect on NO synthesis and iNOS mRNA expression in RAW 264.7 cells. In the case of lipopolysaccharide (LPS) stimulation, NO production and iNOS mRNA expression were detected in RAW 264.7 cells. However, NO production and iNOS mRNA expression which is induced by LPS decreased after treatment of ARE. These data demonstrate that ARE can reduce the suppression of macrophage cell proliferation induced by MTX, and induce IL-1alpha, IL-1beta and IL-6 mRNA expressions in RAW 264.7 macrophage cells. Also, ARE inhibit NO production in LPS-stimulated RAW 264.7 macrophage cells, and the inhibition of NO production may be associated with the inhibition of iNOS mRNA expression.
Lee
Young Sun
YS
The Chinese Medicine Unit, RMIT University, Bundoora, Vic. 3083, Australia.
Han
Ok Kyung
OK
Park
Chan Woo
CW
Yang
Chae Ha
CH
Jeon
Tae Won
TW
Yoo
Wang Keun
WK
Kim
Seong Ho
SH
Kim
Hyo Jung
HJ
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D007887:Fabaceae / Q000737:chemistry*; D015870:Gene Expression / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000964:Antimetabolites, Antineoplastic; D016207:Cytokines; D008070:Lipopolysaccharides; D010936:Plant Extracts; D012333:RNA, Messenger; D009569:Nitric Oxide; D012313:RNA; D008727:Methotrexate