BackgroundIt has been recently noticed that type 2 diabetes (T2D), one of the most common metabolic diseases, causes a chronic low-grade inflammation and activation of the innate immune system that are closely involved in the pathogenesis of T2D. Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3'-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. The molecular mechanisms of cordycepin in T2D are not clear. In the present study, we tested the role of cordycepin on the anti-diabetic effect and anti-inflammatory cascades in LPS-stimulated RAW 264.7 cells.MethodsWe confirmed the levels of diabetes regulating genes mRNA and protein of cytokines through RT-PCR and western blot analysis and followed by FACS analysis for the surface molecules.ResultsCordycepin inhibited the production of NO and pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α in LPS-activated macrophages via suppressing protein expression of pro-inflammatory mediators. T2D regulating genes such as 11β-HSD1 and PPARγ were decreased as well as expression of co-stimulatory molecules such as ICAM-1 and B7-1/-2 were also decreased with the increment of its concentration. In accordance with suppressed pro-inflammatory cytokine production lead to inhibition of diabetic regulating genes in activated macrophages. Cordycepin suppressed NF-κB activation in LPS-activated macrophages.ConclusionBased on these observations, cordycepin suppressed T2D regulating genes through the inactivation of NF-κB dependent inflammatory responses and suggesting that cordycepin will provide potential use as an immunomodulatory agent for treating immunological diseases.

BackgroundChronic 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.MethodsWe confirmed the mRNA levels of the M1/M2 cytokine genes through RT-PCR and morphological change.ResultsLPS-activated macrophages returned to their inactivated original shape, i.e., they looked like naïve macrophages, through the treatment with high concentrations of cordycepin (40 µg/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-1β, TNF-α) and chemokines (CX3CR1, RANTES), as well as cordycepin. Additionally, M2 cytokines (IL-10, IL-1ra, TGF-β) were up-regulated by both cordycepin and adenosine.ConclusionBased 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.

BackgroundSalvia miltiorrhiza (SM) has been used to treat inflammatory diseases including edema and arthritis; however, the anti-inflammatory mechanism of SM action remains unresolved.MethodsThe effects of an ethanol extract of SM (ESM) on pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6, and NO, on anti-inflammatory cytokines including IL-4, IL-10, TGF-β, and IL-1Ra have been studied in an attempt to elucidate the anti-inflammatory mechanism in murine macrophages.ResultsESM inhibited the production of pro-inflammatory cytokines via down-regulation of gene and protein expression whereas it increased the anti-inflammatory cytokines. Furthermore, ESM inhibited the expression of the chemokines, RANTES and CX3CL1, as well as of inflammatory mediators such as TLR-4 and 11β-HSD1.ConclusionThese results indicated that the regulatory effects of ESM may be mediated though the suppression of pro-inflammatory cytokines as well as the induction of anti-inflammatory cytokines. Consequently, we speculate that ESM has therapeutic potential for inflammation-associated disorders.

Cordyceps militaris has been used in traditional medicine to treat numerous diseases and reported to have antitumor and immunomodulatory activities in vivo and in vitro. The precise mechanism of its immune response activities is unclear. The water extract of Cordyceps militaris (CME) and cordycepin (3'-deoxyadenosine) were prepared. Cordycepin, nucleoside analogue, is a metabolite of C. militaris, whereas CME is included the cordycepin 1.3 ng/mg. How CME or cordycepin modulated immune reaction mechanism in antigen presenting cells (APCs) by ability to present exogenous antigens in association with the major histocompatibility complexes (MHC) in vitro and ex-vivo were examined. Microencapsulated ovalbumin (OVA) was efficiently captured, processed and presented on both MHC class I as well as MHC class II molecules. DC 2.4 cells (H-2Kb) or bone marrow- drived DCs (BM-DCs) generated from the BM cells of C57BL/6 mouse (H-2Kb) were cultured in the presence of CME or codycepin with OVA-microspheres, and the amount of OVA peptide - MHC class I complexes was measured by T cell hybridoma, CD8 OVA 1.3 T cell, that recognizes the OVA-H-2Kb complex and IL-2 production. The increment of the phagocytotic activity and the expression of MHC molecules on DCs by CME (12.5 ~ 200 μg/ml) were shown, while suppression of its activity had shown by cordycepin. CME also increased IL-2 production when we examined the effects of CME on CD4+ T cells. On the other hand, cordycepin inhibited CD4+ T cell responses in a concentration of between 5μg to 40μg. The present study demonstrates the modulation of cross priming capability could be a target of therapeutic immunoregulation. Key words: Cordyceps militaris, cross presentation, immunomodulator

Salvia miltiorrhiza (SM) has been commonly used in traditional medicine to treat vascular diseases such as hypertension, arteriosclerosis, and inflammatory disease. It is little kwon that extract of SM alters the immune response of inflammatory reaction. Macrophage, essential cellular components of the body's host defense system, has critical functions in both innate and adoptive system. Macrophages exhibit a particularly vigorous response to endotoxin, which induces production of a variety of inflammatory mediators, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), prostagrandinE2 (PGE2), and nitric oxide (NO) via the inducible nitric oxide synthase (iNOS). In this study, we examined that inhibition effect of ethanol extract of SM (ESM) on pro-inflammatory mediators in LPS-induced murine macrophage cell line RAW 264.7 cells as well as in primary macrophages. ESM deceased the production NO and pro-inflammatory cytokine such as IL-1β, IL-6, TNF-α, and COX-2 in LPS-stimulated macrophage. The variation of gene expression level by ESM treatment was determined by RT-PCR and the relation of mRNA expression of pro-inflammatory cytokine in activated macrophage was suppressed by ESM. In western blot analysis, the level of cytokine protein was decreased in activated macrophage by ESM dose-dependently manner. In addition, the expression of co-stimulatory molecules such as ICAM-1, B7-1, and B7-2 was decreased by ESM dose-dependently. These results showed that ESM has immunomodulatory activities in both innate and adoptive immunity, and they provide the critical information for the understanding of the mechanism of inflammation studies. Key words: Salvia miltiorrhiza, anti-inflammatory, immunomodulator, cytokines