不同真菌固体发酵对蕨菜主要活性成分及其体外抗氧化和抗炎症作用的影响
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摘要
以蕨菜为原料,采用木蹄层孔菌、灵芝和裂褶菌3种功能真菌进行固体发酵,研究发酵前后蕨菜醇提物中总酚、总黄酮等活性成分的变化,并对发酵前后蕨菜体外抗氧化、抗炎症活性进行评价。结果表明,3种真菌固体发酵后蕨菜醇提物的总酚含量较发酵前明显下降,而发酵后总黄酮含量较发酵前均显著增加(P<0.05)。未发酵蕨菜醇提物清除1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl radical,DPPH)自由基和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐自由基(2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate) radical,ABTS~+·)能力均优于固体发酵后的蕨菜醇提物,其中未发酵蕨菜醇提物清除ABTS~+·作用的半数抑制浓度(half-inhibition concentration,IC_(50))值为(0.51±0.02)mg/mL,而经木蹄层孔菌、灵芝和裂褶菌固体发酵后IC_(50)值分别为(2.69±0.01)、(1.20±0.14)、(0.92±0.01)mg/mL。发酵与未发酵蕨菜醇提物在0～0.2 mg/mL质量浓度条件下,对RAW264.7细胞无毒性作用。发酵与未发酵蕨菜醇提物均能抑制脂多糖(lipopolysaccharides,LPS)诱导的RAW264.7细胞一氧化氮(nitric oxide,NO)的分泌水平,且呈现量效关系,其中3种真菌固体发酵后的抑制效果更显著(P<0.05)。当质量浓度为0.2 mg/m L时,未发酵蕨菜醇提物对LPS诱导的RAW264.7细胞NO释放水平的抑制率为27.09%,而经木蹄层孔菌、灵芝和裂褶菌固体发酵后的抑制率分别为58.61%、63.57%、69.46%。相关性分析表明,发酵前后蕨菜醇提物的总酚含量与其抗氧化能力有较好的相关性,总黄酮含量与抗炎症作用呈显著正相关。
Pteridium aquilinum was subjected to solid-state fermentation with Fomes fomentarius, Ganoderma lucidum or Schizophyllum commune in order to investigate changes in the contents of total phenolics and total flavonoids and antioxidant and anti-inflammatory activities in vitro of the ethanol extract of P. aquilinum before and after the fermentation process. The results showed that the total phenolic and total flavonoid contents in the ethanol extract(PAEE) of P. aquilinum was significantly decreased and increased after fungal fermentation, respectively(P < 0.05). PAEE from non-fermented P. aquilinum had stronger scavenging capacities against 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS) free radicals. The half-maximal inhibition concentration(IC_(50)) of non-fermented PAEE was(0.51 ± 0.02) mg/mL, while it was increased to(2.69 ± 0.01),(1.20 ± 0.14), and(0.92 ± 0.01) mg/mL after fermentation with F. fomentarius, G. lucidum, and S. commune, respectively. Both unfermented and fermented PAEE had no significant cytotoxicity in the concentrations range of 0–0.2 mg/mL. In addition, they were capable of inhibiting the production of nitric oxide(NO) in LPS-stimulated RAW264.7 cells in a dose-dependent manner, and the inhibitory effect of fermented PAEE was significantly stronger than that of non-fermented PAEE(P < 0.05). The inhibition percentage of NO production by non-fermented PAEE was 27.09%, while PAEE from P. aquilinum fermented with F. fomentarius, G. lucidum, and S. commune exhibited higher inhibition percentage of 58.61%, 63.57%, and 69.46%, respectively, at a concentration of 0.2 mg/m L. Total phenolic content demonstrated positive correlations with antioxidant activity for native and fermented PAEE. Moreover, significantly positive correlations were found between anti-inflammatory activity and total flavonoid content.
Pteridium aquilinum was subjected to solid-state fermentation with Fomes fomentarius, Ganoderma lucidum or Schizophyllum commune in order to investigate changes in the contents of total phenolics and total flavonoids and antioxidant and anti-inflammatory activities in vitro of the ethanol extract of P. aquilinum before and after the fermentation process. The results showed that the total phenolic and total flavonoid contents in the ethanol extract(PAEE) of P. aquilinum was significantly decreased and increased after fungal fermentation, respectively(P < 0.05). PAEE from non-fermented P. aquilinum had stronger scavenging capacities against 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS) free radicals. The half-maximal inhibition concentration(IC_(50)) of non-fermented PAEE was(0.51 ± 0.02) mg/mL, while it was increased to(2.69 ± 0.01),(1.20 ± 0.14), and(0.92 ± 0.01) mg/mL after fermentation with F. fomentarius, G. lucidum, and S. commune, respectively. Both unfermented and fermented PAEE had no significant cytotoxicity in the concentrations range of 0–0.2 mg/mL. In addition, they were capable of inhibiting the production of nitric oxide(NO) in LPS-stimulated RAW264.7 cells in a dose-dependent manner, and the inhibitory effect of fermented PAEE was significantly stronger than that of non-fermented PAEE(P < 0.05). The inhibition percentage of NO production by non-fermented PAEE was 27.09%, while PAEE from P. aquilinum fermented with F. fomentarius, G. lucidum, and S. commune exhibited higher inhibition percentage of 58.61%, 63.57%, and 69.46%, respectively, at a concentration of 0.2 mg/m L. Total phenolic content demonstrated positive correlations with antioxidant activity for native and fermented PAEE. Moreover, significantly positive correlations were found between anti-inflammatory activity and total flavonoid content.
引文
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[2]马博,苏仕林,李荣峰.蕨菜化学成分及其生物活性研究进展[J].食品工业科技,2011,32(3):413-416.DOI:10.13386/j.issn1002-0306.2011.03.104.
[3]宋广磊,王奎武,张虹,等.蕨菜多糖的分离纯化单糖组分及免疫调节活性研究[J].药物分析杂志,2016,36(12):2107-2112.DOI:10.16155/j.0254-1793.2016.12.06.
[4]TSUMBU C N,DEBY-DUPONT G,TITS M,et al.Polyphenol content and modulatory activities of some tropical dietary plant extracts on the oxidant activities of neutrophils and myeloperoxidase[J].International Journal of Molecular Sciences,2012,13(1):628-650.DOI:10.3390/ijms13010628.
[5]储维维,张烨,段晓梅,等.大理野生蕨菜总黄酮的提取及抗氧化活性研究[J].食品研究与开发,2016,37(2):52-57.DOI:10.3969/j.issn.1005-6521.2016.02.013.
[6]CHEN N D,CHEN N F,CHEN C W,et al.Separation and structure elucidation of a new homoflavanol derivative from Pteridium aquilinum(L.)Kuhn[J].Natural Product Research,2013,27(19):1764-1769.DOI:10.1080/14786419.2012.761618.
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[11]刘东,韩邦兴,胡贺杰,等.蕨菜黄酮提取工艺与药理作用研究进展[J].皖西学院学报,2014,30(2):80-82.DOI:1009-9735(2014)02-0080-03.
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[13]张平,李晔,陈婵,等.灵芝菌株固体发酵对枇杷叶主要活性成分的影响[J].天然产物研究与开发,2016,28(2):242-246.DOI:10.16333/j.1001-6880.2016.2.013.
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[15]SON H U,LEE S,HEO J C,et al.The solid-state fermentation of Artemisia capillaris leaves with Ganoderma lucidum enhances the anti-inflammatory effects in a model of atopic dermatitis[J].International Journal of Molecular Medicine,2017,39(5):1233-1241.DOI:10.3892/ijmm.2017.2945.
[16]KIM H,SUH H J,KANG C M,et al.Immunological activity of ginseng is enhanced by solid-state culture with Ganoderma lucidum mycelium[J].Journal of Medicinal Food,2014,17(1):150-160.DOI:10.1089/jmf.2013.3063.
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[18]邬建国,葛娟,余伟,等.裂褶菌固体发酵葛根渣获取功能产品的条件优化[J].食品工业科技,2010,31(12):192-194.DOI:10.13386/j.issn1002-0306.2010.12.075.
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[20]吴永祥,毕淑峰,姜薇,等.桑白皮多酚对B16细胞内黑色素生成的影响及其机制[J].中国药理学通报,2018,34(9):1296-1301.DOI:10.3969/j.issn.1001-1978.2018.09.021.
[21]李晓英,薛梅,樊汶樵.蓝莓花、茎、叶酚类物质含量及抗氧化活性比较[J].食品科学,2017,38(3):142-147.DOI:10.7506/spkx1002-6630-201703024.
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[28]吴彩琴,陈野,郝迎.固体发酵生产冬虫夏草多糖的研究[J].食品科学,2009,30(7):186-189.DOI:10.3321/j.issn:1002-6630.2009.07.042.
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