鹿茸菇多糖抗氧化保肝研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-0891

摘要/Abstract

摘要：

鹿茸菇是一种珍贵的食药用真菌,鹿茸菇子实体粗多糖具有抗肿瘤、保肝等多种生理活性。以自由基清除率为指标,分析鹿茸菇多糖的化学抗氧化活性,再以H₂O₂建立LO2肝细胞损伤模型,进一步研究鹿茸菇子实体多糖对肝细胞氧化应激损伤的改善机制。结果表明:在体外抗氧化方面,鹿茸菇子实体多糖对DPPH、ABTS、OH自由基具有一定的清除能力,其EC₅₀值分别为:0.218 mg/mL、0.027 mg/mL和2.744 mg/mL;在改善肝细胞氧化应激损伤方面,随着多糖浓度升高,细胞存活率也显著提高,当多糖浓度为5 mmol/mL时,细胞存活率达到94.1%;在清除机体过氧化机制上,相对于模型组,鹿茸菇多糖组抗氧化酶SOD和CAT活力分别提高2.55倍和2.17倍,显著降低细胞内ROS水平,从细胞水平表明鹿茸菇子实体多糖对肝细胞氧化损伤具有一定的改善作用,为鹿茸菇制备保肝类产品提供了一定的研究思路。

关键词: 鹿茸菇, 多糖, 抗氧化

Abstract:

Lyophyllum decastes is a precious edible and medicinal fungus. The crude polysaccharide of Lyophyllum decastes fruit body has many physiological activities such as anti-tumor and liver protection. The free radical scavenging rate was used to analyze the chemical antioxidant activity of L. decastes polysaccharides,and then H₂O₂ was used to establish the LO2 liver cell injury model,and to further study the improvement mechanism of L. decastes fruit body polysaccharides on liver cell oxidative stress damage. The results showed that the polysaccharides from L. decastes fruit body had certain scavenging ability to DPPH,ABTS and OH free radicals in terms of anti-oxidation in vitro,and their EC₅₀ values were:0.218,0.027 and 2.744 mg/mL,respectively. In terms of improving liver cell oxidative stress damage,the cell survival rate also increased significantly as the concentration of polysaccharides increased. The cell survival rate reached 94.1% when the concentration of polysaccharides was 5 mmol/mL. In terms of the mechanism of eliminating the body’s peroxidation,the activities of antioxidant enzymes SOD and CAT increased by 2.55 times and 2.17 times in LDFP group,respectively,compared with the model group,and significantly reduced the level of ROS in cells. The polysaccharide from the fruit body of L. decastes has a certain improvement effect on the oxidative damage of liver cells and provides certain research ideas for the preparation of liver-protecting products of L. decastes.

Key words: Lyophyllum decastes, polysaccharide, antioxidant

张凤培, 徐慧, 邱绍峰, 张君丽, 吴小平, 傅俊生. 鹿茸菇多糖抗氧化保肝研究[J]. 生物技术通报, 2021, 37(11): 92-100.

ZHANG Feng-pei, XU Hui, QIU Shao-feng, ZHANG Jun-li, WU Xiao-ping, FU Jun-sheng. Study on Antioxidant and Liver Protection of Polysaccharide from Lyophyllum decastes[J]. Biotechnology Bulletin, 2021, 37(11): 92-100.

图/表 8

图1 鹿茸菇多糖红外光谱图

Fig.1 Infrared spectrum analysis chart of LDFP

图2 鹿茸菇多糖清除自由基能力 A:OH自由基活力的影响;B:DPPH自由基活力的影响;C:ABTS自由基活力的影响;Vc为阳性对照

Fig. 2 Ability of LDFP to scavenge free radicals A:OH radical scavenging ability. B:DPPH free radical scavenging ability. C:ABTS radical scavenging ability. Vc is positive control

图3 不同浓度鹿茸菇多糖对细胞生长的影响 A:不同质量浓度的多糖对肝细胞LO2的影响;B:不同质量浓度的多糖对肝癌细胞HepG-2的影响。与空白组对比,#:P<0.05,##:P<0.01

Fig. 3 Effect of LDFP in different concentration on the cell growth A:Effects of different concentration of polysaccharides on LO2 cell growth. B:Effects of different concentration of polysaccharides on Hepg-2 cell growth. Compared with the blank group,#:P<0.05;##:P<0.01

图4 LO2细胞氧化损伤模型的建立 A:不同浓度 H2O2 对 LO2 细胞相对存活率的影响. 与空白组对比,#:P<0.05,##:P<0.01;B:不同浓度 H2O2 对人肝细胞 LO2 形态的影响（a:对照组;b:0.05 mmol/L;c:1mmol/L;d:8 mmol/L）

Fig. 4 Establishment of LO2 cell oxidative damage model A:The effect of different concentrations of H2O2 on the relative survival rate of LO2 cells. Compared with the blank group,#:P<0.05;##:P<0.01. B:The effect of different concentrations of H2O2 on the morphology of LO2 cells （a:control group;b:0.05 mmol/L;c:1 mmol/L;d:8 mmol/L）

图5 不同浓度鹿茸菇多糖对氧化损伤的保护作用与空白组对比,#:P<0.05,##:P<0.01;与模型组相比,*:P<0.05,**:P<0.01;下同

Fig. 5 Protective effect of LDFP at different concentrations on oxidative damage Compared with the blank group,#:P<0.05;##:P<0.01. Compared with model group,*:P<0.05;**:P<0.01;the same below

图6 鹿茸菇多糖对肝细胞氧化损伤产生ROS的清除能力

Fig. 6 Ability of LDFP to scavenge ROS produced by oxidative damage of liver cells

图7 鹿茸菇多糖对细胞内SOD酶活表达的调控

Fig. 7 Regulation of LDFP on the expression of SOD enzyme activity in cells

图8 鹿茸菇多糖对细胞内CAT酶活表达的调控

Fig. 8 Regulation of LDFP on the expression of CAT enzyme activity in cells

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