摘要
本试验以浙江省道地药材“浙八味”之一的杭白菊为材料,对其提取液进行体外抗氧化试验,通过抗氧化模型来考查不同诱导剂诱导处理对杭白菊抗氧化活性的影响,从而筛选出其中最佳的诱导剂和最佳诱导方法,并进一步考查了该诱导剂对杭白菊抗氧化防御系统和黄酮类物质合成积累的影响。主要研究结论如下：
1.以杭白菊叶提取物的总还原能力、清除·OH能力和清除DPPH (?).力等为指标,考查了水杨酸、壳聚糖等不同诱导子对杭白菊叶抗氧化活性的影响。结果表明,酵母细胞壁多糖对杭白菊叶抗氧化能力的诱导效果最佳。其中经200mg/L酵母细胞壁多糖处理24h后,总还原能力、清除·OH能力和·DPPH清除率分别可达26.69U/mL、87.12U/mL、77.23%,分别约为对照的1.91、1.39、1.23倍。
2.经酵母细胞壁多糖诱导处理后,杭白菊叶的总酚、总黄酮含量显著提高,说明酵母细胞壁多糖对杭白菊叶酚类和黄酮类物质的合成积累具有促进作用。
3.经500mg/L的酵母细胞壁多糖诱导处理后的24h,杭白菊总抗氧化能力与总黄酮含量之间具有很好的相关性(R2=0.9278),提示酵母细胞壁多糖可能是通过提高杭白菊黄酮类物质合成积累从而提高杭白菊总抗氧化能力。
4.经酵母细胞壁多糖处理后,杭白菊叶中POD的活性大大提高,并且始终高于对照组。诱导后POD活性呈上升趋势,在诱导后的48h达到最大值。杭白菊叶中CAT活性在诱导后的24h达到最大值,其活性约为对照组的1.3倍。经酵母细胞壁多糖诱导后的24h内,处理组杭白菊叶的PPO活性明显高于对照组。上述试验结果表明,酵母细胞壁多糖处理可以激活杭白菊叶中的抗氧化酶系统。
5.经酵母细胞壁多糖诱导处理后,处理组的花和叶中的绿原酸、木犀草苷和蒙花苷这三种主要活性成分含量均明显比对照组高。实验结果表明,酵母细胞壁多糖对杭白菊花和叶中的绿原酸、木犀草苷和蒙花苷这三种主要活性成分的合成积累具有促进作用。
6.酵母细胞壁多糖处理可以显著提高杭白菊叶中黄酮类物质次生代谢合成途径关键酶PAL、C4H活性,表明酵母细胞壁多糖能够激活杭白菊叶中黄酮类物质次生代谢合成途径。因此,推测酵母细胞壁多糖可能通过激活杭白菊叶中黄酮类物质次生代谢合成途径,促进黄酮类物质合成积累。
综上所述,酵母细胞壁多糖处理可以提高杭白菊的抗氧化活性,对杭白菊活性成分的合成有一定的促进作用。提示酵母细胞壁多糖作为一种新型绿色环保的天然调控剂,在药用植物次生代谢产物调控中具有广阔的应用前景。
In this study, Chrysanthemum morifolium extracts were tested in vitro antioxidant tests. In order to filter out the best elicitor and the best induction method, we evaluated the effects of different elicitors on antioxidant activity of Chrysanthemum morifolium through this antioxidant model. Fuether work was to evaluate the effects of this elicitor on antioxidant system and biosynthesis of flavonoids in Chrysanthemum morifolium. The main original research results were shown as follows:
1. After teated with different elicitors, Leaves of Chrysanthemum morifolium were collected for the determination of total reductive capacity,-OH rdical-scavenging activity and-DPPH radical-scavenging activity respectively. Results indicated that antioxidant activities in leaves of Chrysanthemum morifolium treated with yeast cell wall polysaccharides were significantly increased as compared with those of control. In particular, At the24h after treatment of200mg/L yeast cell wall polysaccharides, total reductive capcity,-OH radical-scavenging activity and-DPPH radical-scavenging activity of the leaves were increased to26.69U/mL,87.12U/mL and77.23%respectively, being about1.91-fold,1.39-fold and1.23-fold of control.
2. After yeast cell wall paccharides treatment, total phenols and total flavonoids contents in Chrysanthemum morifolium leaves were significantly increased. It indicated that yeast cell wall polysaccharides had a positive effect on synthesis of phenols and flavonoids in Chrysanthemum morifolium leaves.
3. There was a positive correlation (R2=0.9278) between total flavonoid concentration and total antioxidant activity after24h yeast cell wall polysaccharides (500mg/L) treatment. From the results, we speculated that yeast cell wall polysaccharides may increased total antioxidant capacity by increasing concentration of flavonoids in Chrysanthemum morifolium.
4. After treatment by yeast cell wall polysaccharides, POD activity in Chrysanthemum morifolium leaves greatly increased, and were always higher than control. POD activity reached the maximum at the48h after treatment. At the24h after treatment, CAT activity in Chrysanthemum morifolium leaves reached the maximum, about1.3-fold of control. Within24h after treatment, PPO activity in Chrysanthemum morifolium leaves was significantly higher than control. The results showed that yeast cell wall polysaccharides coud activate antioxidant enzymes system in Chrysanthemum morifolium leaves.
5. Concentrations of chlorogenic acid, luteolin and Linarin in flowers and leaves of Chrysanthemum morifolium were significantly higher than control after treatment by yeast cell wall polysaccharides. It indicated that yeast cell wall polysaccharides had a positive effect on synthesis of chlorogenic acid, luteolin and Linarin in flowers and leaves of Chrysanthemum morifolium.
6. Yeast cell wall polysaccharides can significantly improve the key enzyme PAL, C4H activities of flavonoids secondary metabolic pathway in leaves of Chrysanthemum morifolium, indicating that yeast cell wall polysaccharides could activate flavonoids secondary metabolic pathway in leaves of Chrysanthemum morifolium. So, it was presumed that yeast cell wall polysaccharides promoted the accumulation of flavonoids synthesis through activating flavonoids secondary metabolic pathway in leaves of Chrysanthemum morifolium.
In summary, yeast cell wall polysaccharides could increase the antioxidant activity of Chrysanthemum morifolium, and had a positive effect on synthesis of the active ingredients of Chrysanthemum morifolium. Therefore, yeast cell wall polysaccharides had a broad prospect to be developed as a new type of green natural elicitor used in secondary metabolites regulation of medicinal plants.
引文
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