Immune Netw.  2007 Jun;7(2):57-65. 10.4110/in.2007.7.2.57.

Activation of Macrophages by the Components Produced from Cordyceps militaris

• Kim H
• Kim K
• Han S
• Lee S
• Kwon J
• Lee S
• Kim K

• KMID: 2150627
• DOI: http://doi.org/10.4110/in.2007.7.2.57

Abstract

BACKGROUND: Cordyceps militaris have been reported to modify the immune and inflammatory responses both in vivo and in vitro. Macrophages play important roles in the innate immunity through the phagocytosis of antigens. This study examined the effects of Cordyceps militaris on the activation of murine macrophage RAW 264.7 cells and primary macrophages.
METHODS
The components contained in culture broth of Cordyceps militaris were purified by propyl alcohol extraction and HP 20 column chromatography to CMDB, CMDBW, CMDB5P, and CMDB25P. The amounts of nitric oxide (NO) were determined by using ELISA, Griess reagent respectively. The amounts of some cytokines were determined by using ELISA, western blot, and RT-PCR. The expression levels of cell surface molecules (ICAM-1, B7-1 and B7-2) were measured by flow cytometric analysis.
RESULTS
All the components of Cordyceps militaris produced significant amounts of NO. In particular, CMDB produced much more NO in RAW 264.7 cells and primary macrophages than other fractions of Cordyceps militaris. CMDB increased significantly the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 dose-dependently in RAW 264.7 cells. Examination of the gene expression level also showed that the enhanced production of cytokines was correlated with the up-regulation of i-NOS expression, cycloxygenase (COX)-2 expression, IL-1beta and IL-6 expression, and TNF-alpha expression on the expression of mRNAs by semi-quantitative RT-PCR. Western blot analysis also confirmed that CMDB enhances the expression level of these cytokines.
CONCLUSION
These results show that CMDB stimulates the production of NO and pro-inflammatory cytokines and can also up-regulate the gene expression levels in macrophages.

Keyword

Cordyceps militaris; macrophage; nitric oxide; IL-1beta; TNF-alpha; IL-6

MeSH Terms

1-Propanol
Blotting, Western
Chromatography
Cordyceps*
Cytokines
Enzyme-Linked Immunosorbent Assay
Gene Expression
Immunity, Innate
Interleukin-6
Interleukins
Macrophages*
Nitric Oxide
Phagocytosis
RNA, Messenger
Tumor Necrosis Factor-alpha
Up-Regulation
1-Propanol
Cytokines
Interleukin-6
Interleukins
Nitric Oxide
RNA, Messenger
Tumor Necrosis Factor-alpha