Whitening, Regenerative and Anti－inflammatory Properties of Medicinal Mushroom and Secretion of Snail

• Main Authors: Yu-Ming Wang, 王裕民
• Other Authors: Jane-Yii Wu
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/abdd49

碩士 === 大葉大學 === 生物產業科技學系 === 102 === In the present study, the anti-inflammatory activity of different strains of Cordyceps militaris (CMS, CMW and CMH) and its active components was evaluated via in anti-inflammatory mechanism in RAW 264.7 murine macrophage cells. The results of cytotoxic assay demonstrated that both the fruting body and base body extract did not affect RAW 264.7 cell viability at concentrations up to 125 μg/mL. The water extract of CMS diminished the production of nitric oxide (NO), tumor necrosis factor (TNF)α, and IL-10, in lipopolysaccharide (LPS)-activated RAW264.7 cells and peritoneal macrophages in a dose-dependent manner. CMS also blocked protein expression of inducible NO synthase (iNOS), cyclooxygenase (COX)-2, and TNF-α by suppressing the activation of nuclear factor (NF)-κB. In water extract of CMS, CMW and CMH from fruting body and base body, we also found that uracil, adenine, adenosine and cordycepin are included in. Therefore, we are focus on the evaluation of the individual components of Cordyceps militaris acting as inhibitors of inflammatory. In our results, the uracil and adenosine inhibited the nitrite formation induction in LPS-stimulated RWA264.7 macrophages. Uracil and adenosine also inhibits iNOS and COX-2 expression by blocking NF-κB activation. Furthermore, uracil and adenosine inhibited the IL-10 formation, but had no detectable effect on IL-6 and TNF-α production compared with that of LPS-treated alone cells. In addition, cordycepin inhibited the IL-6 and TNF-α formation, but had no detectable effect on IL-10 production. On the other hand, the mechanism of anti-inflammatory and anti-melanogenic effect of the water extract of Antrodia cinnamomea AC6 grown on grains were investigated. In the effect of Ac-Pb, Ac-Br, Ac-Wt and Ac-Ol on B16-F10 melanoma cell and macrophages cell growth, at 5 mg/mL of Ac-Pb, Ac-Br, Ac-Wt and Ac-Ol, the cell viability was higher than 80%. At 5 mg/mL of Ac-Pb, Ac-Br, Ac-Wt and Ac-Ol, the expressions of COX-2 and iNOS were significantly decreased in RAW 264.7 cell. In the same concentration of Ac-Pb, Ac-Br, Ac-Wt and Ac-Ol markedly inhibited the NO production, IL-6 secretion in dose-dependent manners in LPS-activated macrophage. Moreover, At 5 mg/mL of Ac-Wt and Ac-Ol suppressed TNF-α production and activation of NF-κB. On the other hand, treatment with Ac-Pb, Ac-Br, Ac-Wt and Ac-Ol demonstrated anti-tyrosinase and anti-melanogenic activities in B16 cells. In addtition, we found that either UVA or UVB co-treatment with Ac-Wt, Ac-Br, Ac-Ol and Ac-Pb had inhibitory effect on tyrosinase activity and melanin content, the tyrosinase activity and the melanin content was decreased. In murine B16 melanoma cells stimulated with α-melanocyte-stimulating hormone (α-MSH), Ac-Wt, Ac-Br, Ac-Ol and Ac-Pb not only reduced the melanin content of the cells but also down-regulated tyrosinase activity. On the other hand, the western blotting results showed that Ac-Wt, Ac-Br and Ac-Ol treatment at 5 mg/mL for 24 h significantly reduced Microphthalmia-associated transcription factor (MITF). Furthermore, the focus of the study is possible use of mucus released by Helix aspersa muller (HASL) - a compound inducing cell proliferation. Fibroblasts and macrophages cells treated with HASL at concentrations of 100 μg/mL developed high rates of proliferation, as evaluated using MTT assay and LIVE/DEAD® Cell Viability Assays. In addition, HASL-induced changes on fibroblasts migration were studied by wound-healing assays. On the other hand, At 200-1200 μg/mL of HASL significantly inhibited the LPS-induced NO production. At 100-2000 μg/mL of HASL had no detectable effect on IL-10 production compared with that of LPS-treated alone cells. In conclusion, verification of therapeutic efficacy of fruting body of CMS, Ac-Wt and Ac-Ol as a potent anti-inflammatory remedy. Furthermore, These results shed light on the pharmacological and mechanistic basis of the skin whitening application of Ac-Wt, Ac-Br and Ac-Ol. These results shed light on the regenerative properties of HASL, based on its promoting effect on skin cell migration, proliferation and survival. Moreover, these results support future clinical uses of HASL in the regeneration of wounded tissues.