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      Anti-Inflammatory Properties of the Medicinal Mushroom Cordyceps militaris Might Be Related to Its Linear (1→3)-β-D-Glucan

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          Abstract

          The Ascomycete Cordyceps militaris, an entomopathogenic fungus, is one of the most important traditional Chinese medicines. Studies related to its pharmacological properties suggest that this mushroom can exert interesting biological activities. Aqueous (CW and HW) and alkaline (K5) extracts containing polysaccharides were prepared from this mushroom, and a β-D-glucan was purified. This polymer was analysed by GC-MS and NMR spectrometry, showing a linear chain composed of β-D-Glc p (1→3)-linked. The six main signals in the 13C-NMR spectrum were assigned by comparison to reported data. The aqueous (CW, HW) extracts stimulated the expression of IL-1β, TNF-α, and COX-2 by THP-1 macrophages, while the alkaline (K5) extract did not show any effect. However, when the extracts were added to the cells in the presence of LPS, K5 showed the highest inhibition of the pro-inflammatory genes expression. This inhibitory effect was also observed for the purified β-(1→3)-D-glucan, that seems to be the most potent anti-inflammatory compound present in the polysaccharide extracts of C. militaris. In vivo, β-(1→3)-D-glucan also inhibited significantly the inflammatory phase of formalin-induced nociceptive response, and, in addition, it reduced the migration of total leukocytes but not the neutrophils induced by LPS. In conclusion, this study clearly demonstrates the anti-inflammatory effect of β-(1→3)-D-glucan.

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          Most cited references38

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          Ethical guidelines for investigations of experimental pain in conscious animals.

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            Proinflammatory cytokines.

            To review the concept of proinflammatory cytokines. Review of published literature. Academic (university hospital). Cytokines are regulators of host responses to infection, immune responses, inflammation, and trauma. Some cytokines act to make disease worse (proinflammatory), whereas others serve to reduce inflammation and promote healing (anti-inflammatory). Attention also has focused on blocking cytokines, which are harmful to the host, particularly during overwhelming infection. Interleukin (IL)-1 and tumor necrosis factor (TNF) are proinflammatory cytokines, and when they are administered to humans, they produce fever, inflammation, tissue destruction, and, in some cases, shock and death. Reducing the biological activities of IL-1 and TNF is accomplished by several different, but highly specific, strategies, which involve neutralizing antibodies, soluble receptors, receptor antagonist, and inhibitors of proteases that convert inactive precursors to active, mature molecules. Blocking IL-1 or TNF has been highly successful in patients with rheumatoid arthritis, inflammatory bowel disease, or graft-vs-host disease but distinctly has not been successful in humans with sepsis. Agents such as TNF-neutralizing antibodies, soluble TNF receptors, and IL-1 receptor antagonist have been infused into > 10,000 patients in double-blind, placebo-controlled trials. Although there has been a highly consistent small increase (2 to 3%) in 28-day survival rates with anticytokine therapy, the effect has not been statistically significant. Anticytokine therapy should be able to "rescue" the patient whose condition continues to deteriorate in the face of considerable support efforts. Unfortunately, it remains difficult to identify those patients who would benefit from anticytokine therapy for septic shock.
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              The formalin test in mice: dissociation between inflammatory and non-inflammatory pain.

              The formalin test in mice is a valid and reliable model of nociception and is sensitive for various classes of analgesic drugs. The noxious stimulus is an injection of dilute formalin (1% in saline) under the skin of the dorsal surface of the right hindpaw. The response is the amount of time the animals spend licking the injected paw. Two distinct periods of high licking activity can be identified, an early phase lasting the first 5 min and a late phase lasting from 20 to 30 min after the injection of formalin. In order to elucidate the involvement of inflammatory processes in the two phases, we tested different classes of drugs in the two phases independently. Morphine, codeine, nefopam, and orphenadrine, as examples of centrally acting analgesics, were antinociceptive in both phases. In contrast, the non-steroid anti-inflammatory drugs indomethacin and naproxen and the steroids dexamethasone and hydrocortisone inhibited only the late phase, while acetylsalicylic acid (ASA) and paracetamol were antinociceptive in both phases. The results demonstrate that the two phases in the formalin test may have different nociceptive mechanisms. It is suggested that the early phase is due to a direct effect on nociceptors and that prostaglandins do not play an important role during this phase. The late phase seems to be an inflammatory response with inflammatory pain that can be inhibited by anti-inflammatory drugs. ASA and paracetamol seem to have actions independent of their inhibition of prostaglandin synthesis and they also have effects on non-inflammatory pain.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                17 October 2014
                : 9
                : 10
                : e110266
                Affiliations
                [1 ]Department of Biochemistry and Molecular Biology, Federal University of Parana, Curitiba, Parana, Brazil
                [2 ]Department of Pharmacology, Federal University of Parana, Curitiba, Parana, Brazil
                [3 ]Plant Research International, Wageningen University and Research Centre, Wageningen, The Netherlands
                Shanghai University of Traditional Chinese Medicine, China
                Author notes

                Competing Interests: The authors have read the journal's policy and the authors of this manuscript have the following competing interests: LVG is a retired professor who owns a small non-profit company that provides for reagents and for rent of laboratory and office space. LVG is not involved in any commercial consulting or production related activities concerning Cordyceps sp. The authors confirm that this does not alter their adherence to all PLOS ONE policies on sharing data and materials. There are no restrictions on sharing of data and/or materials.

                Conceived and designed the experiments: FRS CHB DGB APSF GLS MI LVG. Performed the experiments: FRS CHB DGB APSF. Analyzed the data: FRS CHB DGB APSF GLS MI LVG. Contributed reagents/materials/analysis tools: FRS CHB DGB APSF GLS MI LVG. Contributed to the writing of the manuscript: FRS CHB DGB APSF GLS MI LVG.

                Article
                PONE-D-14-26896
                10.1371/journal.pone.0110266
                4201515
                25330371
                43f2c49c-0110-46f2-89d3-72246bcf91a9
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 16 June 2014
                : 11 September 2014
                Page count
                Pages: 11
                Funding
                This work was supported by Conselho Naçional de Desenvolvimento Cientifico e Technológico (CNPq) for the postdoctoral scholarship of FRS; Fundaçäo Araucária; and Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior (CAPES) for the doctoral scholarship of CHB. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. LVG received no special funding for this work.
                Categories
                Research Article
                Biology and Life Sciences
                Biochemistry
                Immunology
                Immune Response
                Immunomodulation
                Molecular Biology
                Macromolecular Structure Analysis
                Mycology
                Fungal Biochemistry
                Engineering and Technology
                Industrial Engineering
                Industrial Processes
                Separation Processes
                Fractionation
                Subcellular Fractionation
                Medicine and health sciences
                Complementary and alternative medicine
                Traditional medicine
                Traditional Chinese medicine
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. The relevant data are contained in the body of the paper. Additional cytotoxicity data are supplied in Figure S1.

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                Uncategorized

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