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. 2011 May 16;10:52.
doi: 10.1186/1475-2891-10-52.

Anti-inflammatory Activity of Edible Oyster Mushroom Is Mediated Through the Inhibition of NF-κB and AP-1 Signaling

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Free PMC article

Anti-inflammatory Activity of Edible Oyster Mushroom Is Mediated Through the Inhibition of NF-κB and AP-1 Signaling

Andrej Jedinak et al. Nutr J. .
Free PMC article

Abstract

Background: Mushrooms are well recognized for their culinary properties as well as for their potency to enhance immune response. In the present study, we evaluated anti-inflammatory properties of an edible oyster mushroom (Pleurotus ostreatus) in vitro and in vivo.

Methods: RAW264.7 murine macrophage cell line and murine splenocytes were incubated with the oyster mushroom concentrate (OMC, 0-100 μg/ml) in the absence or presence of lipopolysacharide (LPS) or concanavalin A (ConA), respectively. Cell proliferation was determined by MTT assay. Expression of cytokines and proteins was measured by ELISA assay and Western blot analysis, respectively. DNA-binding activity was assayed by the gel-shift analysis. Inflammation in mice was induced by intraperitoneal injection of LPS.

Results: OMC suppressed LPS-induced secretion of tumor necrosis factor-α (TNF-α, interleukin-6 (IL-6), and IL-12p40 from RAW264.7 macrophages. OMC inhibited LPS-induced production of prostaglandin E2 (PGE2) and nitric oxide (NO) through the down-regulation of expression of COX-2 and iNOS, respectively. OMC also inhibited LPS-dependent DNA-binding activity of AP-1 and NF-κB in RAW264.7 cells. Oral administration of OMC markedly suppressed secretion of TNF-α and IL-6 in mice challenged with LPS in vivo. Anti-inflammatory activity of OMC was confirmed by the inhibition of proliferation and secretion of interferon-γ (IFN-γ), IL-2, and IL-6 from concanavalin A (ConA)-stimulated mouse splenocytes.

Conclusions: Our study suggests that oyster mushroom possesses anti-inflammatory activities and could be considered a dietary agent against inflammation. The health benefits of the oyster mushroom warrant further clinical studies.

Figures

Figure 1
Figure 1
Effect of OMC on viability of macrophages. RAW264.7 cells were treated with OMC (0-100 μg/ml) for 24, 48 and 72 hours and cell proliferation-viability determined as described in Materials and Methods. The data are means ± S.D., n = 3-4.
Figure 2
Figure 2
Effect of OMC on LPS-induced PGE2 and NO secretion and COX-2 and iNOS expression in RAW264.7 cells. (A) PGE2 and (C) NO secretion were determined in cell culture media from RAW264.7 cells treated with OMC and LPS as described in Materials and Methods. The data are means ± S.D. of two independent experiments, repeated minimally twice. Means without a common letter differ, P < 0.05. Expression of (B) COX-2 and (D) iNOS were determined in whole cell lysates from RAW264.7 cells treated with OMC and LPS as described in Materials and Methods. The equal protein loading was verified with anti-β-actin antibody. The results are representative of three separate experiments. (E) Secretion of IL-6 was determined in cell culture media from RAW264.7 cells treated with OMC and TNF-α as described in Materials and Methods. The data are means ± S.D. of two independent experiments, repeated minimally twice. Means without a common letter differ, P < 0.05.
Figure 3
Figure 3
Effect of OMC on LPS-dependent activation of AP-1, NF-κB and STAT3 in RAW264.7 cells. (A) AP-1 and (B) NF-κB DNA-binding activity was determined by gel shift analysis in nuclear extracts isolated from RAW264.7 cells pretreated with OMC (0, 50 100 μg/ml) for 24 hours followed by the stimulation with LPS (1 μg/ml) for an additional 30 minutes. Nuclear extracts were subjected to EMSA with a [32P]-labeled AP-1 or [32P]-labeled NF-κB probe as described in Materials and Methods. The specificity of DNA-binding was confirmed by competitive gel shift with cold AP-1, NF-κB, or unrelated DNA (URL). (C) STAT3 activity was evaluated in whole cell extracts treated with OMC for 24 hours and LPS for 30 minutes by Western blot analysis with anti-phospho-STAT3 antibody. The equal protein loading was verified with anti-STAT3 antibody. The results are representative of three separate experiments.
Figure 4
Figure 4
OMC reduces LPS-induced cytokine production in vivo. (A) TNF-α and (B) IL-6 were determined in plasma in mice treated with OMC (1000 mg/kg) and LPS (0.1 mg/kg) as described in Materials and Methods. The data are means ± S.D, n = 3-6. Means without a common letter differ, P < 0.05.
Figure 5
Figure 5
OMC inhibits ConA-induced splenocyte proliferation and production of IFN-γ, IL-6 and IL-4. Splenocytes were cultured in the presence of ConA (5 μg/ml) and OMC (0-100 μg/ml) for 72 hours. (A) cell proliferation, (B) production of IFN-γ, (C) IL-2, and (D) IL-4 was determined as described in Materials and Methods. The data are means ± S.D. of triplicate measurements. Similar data were obtained in two additional independent experiments. Means without a common letter differ, P < 0.05.

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