灰树花不同浓度醇提物的活性成分分析及抗氧化、抗EV71病毒的作用研究

熊雯宇; 何君强; 汤丹琳; 彭培植; 江小琴; 刘斌

doi:10.19303/j.issn.1008-0384.2023.04.014

灰树花不同浓度醇提物的活性成分分析及抗氧化、抗EV71病毒的作用研究

Active Ingredients and Antioxidative and Anti-EV71 Virus Effects of Grifola frondosa Ethanol Extract

摘要

摘要:
目的探究灰树花适宜的乙醇提取浓度，分析不同乙醇浓度提取对灰树花活性成分及抗氧化、抗EV71病毒活性的影响，促进灰树花的开发利用。
方法分别采用55%、70%和95%的乙醇溶液作为提取溶剂对灰树花子实体进行超声波辅助提取，测定不同浓度乙醇提取物主要活性成分含量并对其抗氧化、抗EV71病毒能力进行相关性分析。
结果不同乙醇浓度提取物活性成分含量存在差异，灰树花95%乙醇提取物（GFE95）中多酚和三萜含量最高分别为0.042%和1.82%，灰树花55%乙醇提取物（GFE55）中多糖和蛋白质含量最高，分别为33.08%和17.45%。GFE55、灰树花70%乙醇提取物（GFE70）和GFE95具有不同程度的抗氧化和EV71病毒抑制作用。其中，GFE95的抗氧化能力最好，对2,2-联苯基-1-苦基肼基（DPPH）、2,2′-联氨双二胺盐（ABTS⁺）、超氧阴离子（O₂⁻）和过氧化氢（H₂O₂）自由基的清除能力的IC₅₀值分别为0.7427、0.7414、0.902、0.4146 mg·mL⁻¹。GFE95的抗病毒作用较强，当样品质量浓度为200 μg·mL⁻¹时，GFE95对EV71病毒的抑制率最高，达88.18%，其对EV71病毒抑制率的IC₅₀值为194.80 μg·mL⁻¹，TI值为9.65。皮尔逊相关性分析结果表明DPPH、ABTS⁺、O₂⁻自由基清除率与三萜类成分含量呈极显著正相关，相关系数分别为0.992、0.971和0.613（P＜0.01）。O₂⁻、H₂O₂自由基清除率与多酚类成分含量呈极显著正相关，相关系数分别为0.921和0.997（P＜0.01）。EV71病毒抑制作用与多酚、三萜类成分含量呈极显著正相关（P＜0.01），相关系数分别为0.536和0.954。灰树花中三萜和多酚的含量与抗氧化、抗病毒活性具有较强的相关性。
结论以体积分数为95%的乙醇溶液为提取剂，可有效提取灰树花中具有抗氧化、抗病毒活性的多酚、三萜、多糖等活性物质，提高灰树花的生物活性，是进一步促进灰树花的高值化应用开发的关键。研究结果为灰树花的提取溶剂选择及活性作用的研究奠定基础。

Abstract:
Objective Active ingredients and antioxidative and anti-EV71 virus activities of the Grifola frondose extracts using varied concentration of ethanol were analyzed.
Method Using 55%, 70%, and 95% ethanol solutions, a ultrasound-assisted extraction was conducted to produce concoctions from G.frondosa fruiting bodies.Content of active ingredients and efficacy of the extracts obtained with varied concentrations of ethanol as an antioxidant and anti-EV71 virus agent were determined.
Result The content of polyphenols at 0.042% and that of triterpenes at 1.82% were the highest in the 95% ethanol extract (GFE95) of all concoctions.The maximal polysaccharides at 33.08% and proteins at 17.45% were found in GFE55 that used 55% ethanol for the extraction.GFE95 also showed the greatest antioxidant capacity with IC₅₀ on scavenging DPPH, ABTS⁺, O₂⁻, and H₂O₂ free radicals of 0.742 7 mg·mL⁻¹, 0.741 4 mg·mL⁻¹, 0.902 6 mg·mL⁻¹, and 0.414 6 mg·mL⁻¹, respectively, and a strong antiviral effect on EV71 at the concentration of 200 μg·mL⁻¹ with the inhibition rate of 88.18%, an IC₅₀ of 194.80 μg·mL⁻¹, and a TI of 9.65.A Pearson’s correlation analysis indicated that the DPPH, ABTS⁺, and O₂⁻ free radical scavenging rates significantly correlated to the triterpenoids content with correlation coefficients of 0.992, 0.971, and 0.613, respectively (P＜0.01); the O₂⁻ and H₂O₂ free radical scavenging rates significantly correlated to the polyphenols with correlation coefficients of 0.921 and 0.997, respectively (P＜0.01); the anti-EV71 virus effect significantly correlated to the polyphenols and triterpenoids with correlation coefficients of 0.536 and 0.954, respectively (P＜0.01); and the triterpenoids and polyphenols significantly correlated to the antioxidative and antiviral activities of the extract.
Conclusion The 95% ethanol extraction of G.frondosa optimally retrieved the active substances, such as polyphenols, triterpenes, and polysaccharides, in the fruiting bodies.The extract exhibited significant antioxidative and antiviral capacities, with which a valuable pharmaceutical material might be developed.

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