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Attenuation of Inflammatory Mediators (TNF-α and Nitric Oxide) and Up-Regulation of IL-10 by Wild and Domesticated Basidiocarps of Amauroderma rugosum (Blume & T. Nees) Torrend in LPS-Stimulated RAW264.7 Cells.

Chan PM, Tan YS, Chua KH, Sabaratnam V, Kuppusamy UR - PLoS ONE (2015)

Bottom Line: There was no inhibitory effect of WB and DB on nuclear translocation of NF-κB p65.In conclusion, the wild and domesticated basidiocarps of A. rugosum possessed antioxidant and in vitro anti-inflammatory properties.No inhibitory effects shown on upstream nuclear translocation of NF-κB p65.

View Article: PubMed Central - PubMed

Affiliation: Mushroom Research Centre, University of Malaya, 50603, Kuala Lumpur, Malaysia; Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
Amauroderma rugosum, commonly known as "Jiǎzī" in China, is a wild mushroom traditionally used by the Chinese to reduce inflammation, to treat diuretic and upset stomach, and to prevent cancer. It is also used by the indigenous communities in Malaysia to prevent epileptic episodes and incessant crying by babies. The aim of this study was to compare the wild and domesticated basidiocarps of A. rugosum for antioxidant and in vitro anti-inflammatory effects in LPS-stimulated RAW264.7 cells. The wild basidiocarps of A. rugosum were collected from the Belum Forest, Perak, Malaysia and the domesticated basidiocarps of A. rugosum were cultivated in the mushroom house located in the University of Malaya, Kuala Lumpur, Malaysia. Both the wild and domesticated basidiocarps were subjected to ethanolic extraction and the extracts were tested for antioxidant and anti-inflammatory activities. In this study, the crude ethanolic extract of wild (WB) and domesticated (DB) basidiocarps of A. rugosum had comparable total phenolic content and DPPH scavenging activity. However, WB (EC50 = 222.90 μg/mL) displayed a better ABTS cation radical scavenging activity than DB (EC50 = 469.60 μg/mL). Both WB and DB were able to scavenge nitric oxide (NO) radical and suppress the NO production in LPS-stimulated RAW264.7 cells and this effect was mediated through the down-regulation of inducible nitric oxide synthase (iNOS) gene. In addition, both WB and DB caused down-regulation of the inflammatory gene TNF-α and the up-regulation of the anti-inflammatory gene IL-10. There was no inhibitory effect of WB and DB on nuclear translocation of NF-κB p65. In conclusion, the wild and domesticated basidiocarps of A. rugosum possessed antioxidant and in vitro anti-inflammatory properties. WB and DB inhibited downstream inflammatory mediators (TNF-α and NO) and induced anti-inflammatory cytokine IL-10 production. No inhibitory effects shown on upstream nuclear translocation of NF-κB p65. WB and DB exhibited antioxidant activity and attenuation of proinflammatory mediators and therefore, A. rugosum may serve as a potential therapeutic agent in the management of inflammation.

No MeSH data available.


Related in: MedlinePlus

Effects of A. rugosum extracts on LPS-induced NO production by RAW264.7 cells (S1 Dataset).RAW264.7 cells were co-incubated with various concentrations of A. rugosum extracts and 1 μg/mL LPS for 24 hrs. L-NAME (250 μM) served as the positive control. Results shown represent the mean ± SD, n = 3 and *p < 0.05 versus LPS-induced NO level alone.
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pone.0139593.g003: Effects of A. rugosum extracts on LPS-induced NO production by RAW264.7 cells (S1 Dataset).RAW264.7 cells were co-incubated with various concentrations of A. rugosum extracts and 1 μg/mL LPS for 24 hrs. L-NAME (250 μM) served as the positive control. Results shown represent the mean ± SD, n = 3 and *p < 0.05 versus LPS-induced NO level alone.

Mentions: The inhibitory effect of WB and DB on LPS-stimulated NO production in RAW264.7 cells were evaluated using the Griess reaction. Both WB and DB showed inhibition of NO production in LPS-stimulated RAW264.7 cells (Fig 3). At the concentration of 0.1 μg/mL, WB and DB were able to suppress the NO production in LPS-stimulated RAW264.7 cells by 46.79% (14.13 ± 0.69 μM) and 49.03% (13.53 ± 2.07 μM), respectively. L-NAME, a standard NOS inhibitor, used as a positive control in this study significantly (p < 0.001) inhibited NO production (5.91 ± 1.31 μM, 83.50% inhibition) at 250 μM. Fig 2 shows that the inhibitory actions of WB and DB on NO production were not due to cell death. Besides that, the inducible nitric oxide synthase (iNOS) gene in RAW264.7 cells was up-regulated upon stimulation with 1 μg/mL of LPS and was significantly down-regulated with the treatment of WB and DB at the concentration of 0.1 μg/mL (Figs 4, 5A and 5B).


Attenuation of Inflammatory Mediators (TNF-α and Nitric Oxide) and Up-Regulation of IL-10 by Wild and Domesticated Basidiocarps of Amauroderma rugosum (Blume & T. Nees) Torrend in LPS-Stimulated RAW264.7 Cells.

Chan PM, Tan YS, Chua KH, Sabaratnam V, Kuppusamy UR - PLoS ONE (2015)

Effects of A. rugosum extracts on LPS-induced NO production by RAW264.7 cells (S1 Dataset).RAW264.7 cells were co-incubated with various concentrations of A. rugosum extracts and 1 μg/mL LPS for 24 hrs. L-NAME (250 μM) served as the positive control. Results shown represent the mean ± SD, n = 3 and *p < 0.05 versus LPS-induced NO level alone.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591274&req=5

pone.0139593.g003: Effects of A. rugosum extracts on LPS-induced NO production by RAW264.7 cells (S1 Dataset).RAW264.7 cells were co-incubated with various concentrations of A. rugosum extracts and 1 μg/mL LPS for 24 hrs. L-NAME (250 μM) served as the positive control. Results shown represent the mean ± SD, n = 3 and *p < 0.05 versus LPS-induced NO level alone.
Mentions: The inhibitory effect of WB and DB on LPS-stimulated NO production in RAW264.7 cells were evaluated using the Griess reaction. Both WB and DB showed inhibition of NO production in LPS-stimulated RAW264.7 cells (Fig 3). At the concentration of 0.1 μg/mL, WB and DB were able to suppress the NO production in LPS-stimulated RAW264.7 cells by 46.79% (14.13 ± 0.69 μM) and 49.03% (13.53 ± 2.07 μM), respectively. L-NAME, a standard NOS inhibitor, used as a positive control in this study significantly (p < 0.001) inhibited NO production (5.91 ± 1.31 μM, 83.50% inhibition) at 250 μM. Fig 2 shows that the inhibitory actions of WB and DB on NO production were not due to cell death. Besides that, the inducible nitric oxide synthase (iNOS) gene in RAW264.7 cells was up-regulated upon stimulation with 1 μg/mL of LPS and was significantly down-regulated with the treatment of WB and DB at the concentration of 0.1 μg/mL (Figs 4, 5A and 5B).

Bottom Line: There was no inhibitory effect of WB and DB on nuclear translocation of NF-κB p65.In conclusion, the wild and domesticated basidiocarps of A. rugosum possessed antioxidant and in vitro anti-inflammatory properties.No inhibitory effects shown on upstream nuclear translocation of NF-κB p65.

View Article: PubMed Central - PubMed

Affiliation: Mushroom Research Centre, University of Malaya, 50603, Kuala Lumpur, Malaysia; Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.

ABSTRACT
Amauroderma rugosum, commonly known as "Jiǎzī" in China, is a wild mushroom traditionally used by the Chinese to reduce inflammation, to treat diuretic and upset stomach, and to prevent cancer. It is also used by the indigenous communities in Malaysia to prevent epileptic episodes and incessant crying by babies. The aim of this study was to compare the wild and domesticated basidiocarps of A. rugosum for antioxidant and in vitro anti-inflammatory effects in LPS-stimulated RAW264.7 cells. The wild basidiocarps of A. rugosum were collected from the Belum Forest, Perak, Malaysia and the domesticated basidiocarps of A. rugosum were cultivated in the mushroom house located in the University of Malaya, Kuala Lumpur, Malaysia. Both the wild and domesticated basidiocarps were subjected to ethanolic extraction and the extracts were tested for antioxidant and anti-inflammatory activities. In this study, the crude ethanolic extract of wild (WB) and domesticated (DB) basidiocarps of A. rugosum had comparable total phenolic content and DPPH scavenging activity. However, WB (EC50 = 222.90 μg/mL) displayed a better ABTS cation radical scavenging activity than DB (EC50 = 469.60 μg/mL). Both WB and DB were able to scavenge nitric oxide (NO) radical and suppress the NO production in LPS-stimulated RAW264.7 cells and this effect was mediated through the down-regulation of inducible nitric oxide synthase (iNOS) gene. In addition, both WB and DB caused down-regulation of the inflammatory gene TNF-α and the up-regulation of the anti-inflammatory gene IL-10. There was no inhibitory effect of WB and DB on nuclear translocation of NF-κB p65. In conclusion, the wild and domesticated basidiocarps of A. rugosum possessed antioxidant and in vitro anti-inflammatory properties. WB and DB inhibited downstream inflammatory mediators (TNF-α and NO) and induced anti-inflammatory cytokine IL-10 production. No inhibitory effects shown on upstream nuclear translocation of NF-κB p65. WB and DB exhibited antioxidant activity and attenuation of proinflammatory mediators and therefore, A. rugosum may serve as a potential therapeutic agent in the management of inflammation.

No MeSH data available.


Related in: MedlinePlus