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Antioxidant Activity of Shiitake Mycelial Exudates.

机译：香菇菌丝体分泌物的抗氧化活性。

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摘要

Shiitake mushroom (Lentinus edodes), known in China as Xiang-gu, is one of the most valuable medicinal mushrooms, and has been used for thousands of years both as food and medicine. Shiitake mushroom extracts have also been found to have antioxidant properties and their antioxidant ability is positively correlated with their phenolic content. However, thus far, investigation of the antioxidant ability of shiitake mushroom has mainly focused on the fruiting body, and the antioxidant properties of its mycelial exudates are rarely reported.;In this study, exudates (DE) secreted from two shiitake mushroom mycelia (strains 1358 and L5458) were evaluated for their antioxidative properties and phenolic content. 1358DE and L5458DE showed distinct antioxidant activity in different in vitro assays, including scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, hydroxyl radical, superoxide anions and hydrogen peroxide; the ability to chelate ferrous ions; reducing power; hemolysis inhibition activity in rat erythrocyte; and lipid peroxidation inhibition (IC50 values of 1358DE and L5458DE were 3.3 and 132.6; 44.5 and > 1000; 26.9 and 53.7; 153.6 and >175.0; 176.0 and 521.0; 26.7 and 746.4; 47.8 and 736.9; and 3.1 and > 1000 microg/mL, respectively). Their total phenolic content was 237.33 and 24.08 mg gallic acid equivalent (GAE)/g of dry DE, respectively. Overall, these results show that 1358DE generally possesses better antioxidant properties than L5458DE, possibly due to its larger total phenolic content.;1358DE were selected to further investigate for its better antioxidant effect. 1358DE was fractionated using water-solvent partition and two fractions [water soluble fraction (WF) and ethyl acetate fraction (EF)] were obtained. The antioxidant effects of WF were similar to those of the original 1358DE, while EF did not possess any antioxidant activities. The WF was further isolated with polyamide column, which can separated the polyphenols from other components, and two sub-fraction (P-1 and P-2) were obtained. After the WF passing through the polyamide column, carbohydrate content in the sub-fraction 1 (P-1) was significantly increased, while its total phenolic content reduced dramatically, and its antioxidant activity decreased. However, the sub-fraction 2 (P-2) was the opposite. Carbohydrate content in P-2 was significantly reduced, while its total phenolic content increased dramatically, and its antioxidant activity increased. Apparently, carbohydrate contributed little to the antioxidant effect than that of the phenolic compounds as shown from this investigation. These results suggest that the antioxidant effect in 1358DE was contributed by the presence of polyphenols. Besides, results from Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and ferric trichloride reaction suggested that P-2 was oligomers of water soluble polyphenols (WSP) with the molecular weight of about 600∼1200 Da.;The water soluble polyphones (WSP) were the potent antioxidant components in 1358DE and further study its protective effect against the hydrogen peroxide which induced cytotoxicity in V79-4 cells. In the cell viability experiments, pretreatment of WSP at the concentrations of 6.25∼50 microg/ml increased the cell viability significantly more than at the presence of H2O2 only. Besides, the pretreatment of cells with WSP significantly inhibited the increase of Malondialdehyde (MDA, which is a by-product of lipid peroxidation) production and the decrease of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) activities induced by H2O2.;Quite a few antioxidant compounds have been reported that a causative relationship may exist between the anti-angiogenic activity and antioxidant effect. Therefore, a zebrafish model was using to investigate the anti-angiogenic activity of the WSP because of its excellent antioxidant activity. In quantitative of endogenous alkaline phosphatase (EAP) assay, after the embryos treated with WSP at final concentrations of 50, 100, 150, 200, 250 microg/ml, and the vessel formation were significantly (p 0.05) reduced to 87.2, 85.6, 74.8, 69.4, and 62.8% of the control value, respectively. Moreover, from the microscope, compare to the control, WSP at the concentration of 250 microg/ml also showed potent inhibition on the intersegmental blood vessels (ISVs) formation in Tg(fli1a:EGFP)y1 zebrafish embryos. Thus, the finding indicated that WSP could inhibit vessel formation in zebrafish.;Overall, this study revealed that water soluble polyphenols (WSP) was the active components of 1358DE. Besides of the antioxidant effect, the WSP could inhibit vessel formation significantly in zebrafish. The findings indicate that exudates of shiitake mushroom mycelia have good potential as a source of natural antioxidants.
机译：香菇（Lentinus edodes）在中国被称为香菇，是最有价值的药用蘑菇之一，已被用作食品和药品数千年。香菇提取物也被发现具有抗氧化性能，其抗氧化能力与其酚含量呈正相关。然而，到目前为止，香菇抗氧化能力的研究主要集中在子实体上，很少报道其菌丝分泌物的抗氧化特性。;在这项研究中，两个香菇菌丝体（菌株）分泌的分泌物（DE）对1358和L5458）的抗氧化性能和酚含量进行了评估。 1358DE和L5458DE在不同的体外测定法中显示出独特的抗氧化活性，包括对1,1-二苯基-2-吡啶并肼基（DPPH），羟基，超氧阴离子和过氧化氢的清除活性。螯合亚铁离子的能力；降低功率；大鼠红细胞的溶血抑制活性；和脂质过氧化抑制（1358DE和L5458DE的IC50值为3.3和132.6; 44.5和> 1000; 26.9和53.7; 153.6和> 175.0; 176.0和521.0; 26.7和746.4; 47.8和736.9;以及3.1和> 1000 microg / mL ， 分别）。它们的总酚含量分别为237.33和24.08 mg没食子酸当量（GAE）/ g干DE。总体而言，这些结果表明1358DE总体上具有比L5458DE更好的抗氧化性能，这可能是由于其总酚含量较高。选择1358DE来进一步研究其抗氧化效果。使用水分配器将1358DE分馏，得到两个馏分[水溶性馏分（WF）和乙酸乙酯馏分（EF）]。 WF的抗氧化作用与原始的1358DE相似，而EF没有任何抗氧化活性。用聚酰胺柱进一步分离WF，该柱可将多酚与其他组分分离，得到两个亚级分（P-1和P-2）。 WF通过聚酰胺色谱柱后，子级分1（P-1）中的碳水化合物含量显着增加，而其总酚含量急剧下降，其抗氧化活性下降。但是，子分数2（P-2）相反。 P-2中的碳水化合物含量显着降低，而其总酚含量急剧增加，其抗氧化活性增加。显然，如本研究所示，碳水化合物对酚类物质的抗氧化作用几乎不如酚类化合物。这些结果表明，多酚的存在有助于1358DE中的抗氧化作用。此外，基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）和三氯化铁反应的结果表明P-2是水溶性多酚（WSP）的低聚物，分子量约为600〜 1200 Da；水溶性聚醚（WSP）是1358DE中有效的抗氧化剂成分，并进一步研究了其对过氧化氢的保护作用，该过氧化氢可诱导V79-4细胞的细胞毒性。在细胞活力实验中，以6.25〜50 microg / ml的浓度进行WSP预处理比仅存在H2O2时显着增加了细胞活力。此外，用WSP预处理细胞可显着抑制丙二醛（MDA，它是脂质过氧化作用的副产物）的产生，并抑制H2O2诱导的抗氧化酶（超氧化物歧化酶，过氧化氢酶和谷胱甘肽过氧化物酶）活性的降低。已经报道了许多抗氧化剂化合物，其抗血管生成活性和抗氧化剂作用之间可能存在因果关系。因此，斑马鱼模型因其出色的抗氧化活性而被用于研究WSP的抗血管生成活性。在定量的内源性碱性磷酸酶（EAP）分析中，用WSP处理的终浓度为50、100、150、200、250 microg / ml的胚胎和血管形成明显减少（p <0.05）至87.2、85.6分别为对照值的74.8、69.4和62.8％。此外，从显微镜上看，与对照相比，浓度为250微克/毫升的WSP还显示出对Tg（fli1a：EGFP）y1斑马鱼胚胎中节间血管（ISV）形成的有效抑制作用。因此，该发现表明WSP可以抑制斑马鱼的血管形成。总的来说，这项研究表明水溶性多酚（WSP）是1358DE的活性成分。除了抗氧化作用，WSP还可以显着抑制斑马鱼的血管形成。研究结果表明，香菇菌丝体的渗出液具有作为天然抗氧化剂来源的良好潜力。

著录项

  • 作者

    Huang, Weihuan.;

  • 作者单位

    The Chinese University of Hong Kong (Hong Kong).;

  • 授予单位 The Chinese University of Hong Kong (Hong Kong).;
  • 学科 Biology Botany.;Biology Cell.;Health Sciences Nutrition.
  • 学位 Ph.D.
  • 年度 2012
  • 页码 121 p.
  • 总页数 121
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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