Immune Netw.  2009 Dec;9(6):255-264. 10.4110/in.2009.9.6.255.

Role of Cordycepin and Adenosine on the Phenotypic Switch of Macrophages via Induced Anti-inflammatory Cytokines

• Shin S
• Moon S
• Park Y
• Kwon J
• Lee S
• Lee CK
• Cho K
• Ha NJ
• Kim K

• KMID: 2150654
• DOI: http://doi.org/10.4110/in.2009.9.6.255

Abstract

BACKGROUND
Chronic low grade inflammation is closely linked to type II diabetes, obesity, and atherosclerosis. Macrophages play a key role in the regulation of pro- or anti-inflammatory actions at the lesion sites of disease. Components of cordyceps militaris, cordycepin and adenosine, have been used for the modulation of inflammatory diseases. The effects of cordycepin in the modulation of macrophages have yet to be elucidated. We investigated the effects of cordycepin and adenosine on the morphological changes of macrophages under the inflammatory condition of LPS and an anti-inflammatory condition involving high concentrations of adenosine. METHODS: We confirmed the mRNA levels of the M1/M2 cytokine genes through RT-PCR and morphological change. RESULTS: LPS-activated macrophages returned to their inactivated original shape, i.e., they looked like naive macrophages, through the treatment with high concentrations of cordycepin (40 microgram/ml). LPS and adenosine activated macrophages also returned to their original inactivated shapes after cordycepin treatment; however, at relatively higher levels of cordycepin than adenosine. This change did not occur with relatively low concentrations of cordycepin. Adenosine down-regulated the gene expression of M1 cytokines (IL-1beta, TNF-alpha) and chemokines (CX3CR1, RANTES), as well as cordycepin. Additionally, M2 cytokines (IL-10, IL-1ra, TGF-beta) were up-regulated by both cordycepin and adenosine. CONCLUSION: Based on these observations, both cordycepin and adenosine regulated the phenotypic switch on macrophages and suggested that cordycepin and adenosine may potentially be used as immunomodulatory agents in the treatment of inflammatory disease.

Keyword

Cordycepin; Adenosine; M1/M2 phenotype; Morphology change

MeSH Terms

Adenosine
Atherosclerosis
Chemokines
Cordyceps
Cytokines
Deoxyadenosines
Gene Expression
Inflammation
Interleukin 1 Receptor Antagonist Protein
Macrophages
Obesity
RNA, Messenger
Adenosine
Chemokines
Cytokines
Deoxyadenosines
Interleukin 1 Receptor Antagonist Protein
RNA, Messenger

Figure

• 

  Figure 1 RAW 264.7 cells were cultured on cover slips in the presence of various concentrations of cordycepin (A) medium alone (a), LPS 100 ng/ml (b), cordycepin (5 µg/ml) (c), 10 µg/ml (d), 20 µg/ml (e), and 40 µg/ml (f), in the presence of LPS and adenosine (B) medium alone (a), LPS 100 ng/ml (b), adenosine (5 µg/ml) (c), 10 µg/ml (d), 20 µg/ml (e), 40 µg/ml (f), and in the absence of LPS for 24 hrs. The cells were then fixed and stained in Diff-quick Solution (×400).

• 

  Figure 2 Effects of cordycepin and adenosine on the expression of M1 cytokines (A, B), chemokines, and their receptor (C, D) in RAW 264.7 cells. Cells were incubated in various concentrations of cordycepin (5, 10, 20, 40 µg/ml) in the presence of LPS (100 ng/ml) or adenosine (10 µg/ml), in the presence or absence of LPS, for 24 hrs. mRNA levels of M1 cytokine and chemokine genes were determined by RT-PCR analysis. β-actin was used as a control. ††p<0.01 vs. cells only; *p<0.05, **p<0.01 vs. LPS only; and †p<0.05 vs. cells only were all based on Student's t-tests.

• 

  Figure 3 Effects of cordycepin (A) and adenosine (B) on the expression of M2 cytokines in RAW 264.7 cells. Cells were incubated in various concentrations of cordycepin (5, 10, 20, 40 µg/ml) in the presence of LPS (100 ng/ml), or adenosine (10 µg/ml), in the presence or absence of LPS, for 24 hrs. mRNA levels of M2 cytokine genes were determined by RT-PCR analysis. β-actin was used as a control. ††p<0.01 vs. cells only; *p<0.05, **p<0.01 vs. LPS only; and †p<0.01 vs. cells only were based on Student's t-tests.

• 

  Figure 4 The morphological switch by cordycepin with adenosine or adenosine and cordycepin. RAW 264.7 cells were cultured on cover slips in the presence of double the concentration of cordycepin than adenosine (A) in: medium alone (a), LPS 100 ng/ml (b), adenosine (2.5 µg/ml)/cordycepin (5 µg/ml) (c), adenosine (5 µg/ml)/cordycepin (10 µg/ml) (d), adenosine (10 µg/ml)/cordycepin (20 µg/ml) (e), adenosine (20 µg/ml)/cordycepin (40 µg/ml) (f), and 2× adenosine/cordycepin (B) in: medium alone (a), LPS 100 ng/ml (b), adenosine (20 µg/ml)/cordycepin (10 µg/ml) (c), adenosine (40 µg/ml)/cordycepin (20 µg/ml) (d), adenosine (80 µg/ml)/cordycepin (40 µg/ml) (e), for 24 hrs. The cells were then fixed and stained in Diff-quick Solution (×400).

• 

  Figure 5 Lipid accumulation of macrophages by cordycepin. RAW 264.7 cells were cultured on cover slips in the presence of various concentrations of cordycepin: medium alone (A), LPS (100 ng/ml) (B), cordycepin (5 µg/ml) (C), 10 µg/ml (D), 20 µg/ml (E), 40 µg/ml (F) in the presence of LPS, for 24 hrs. The cells were then fixed and stained in oil red O solution. (×400).

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