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This study aimed to optimize the extraction process of Cistanche tubulosa polysaccharides (CP) and investigate their structural characteristics and anti-fatigue effects. Single-factor experiments combined with response surface methodology were employed to optimize extraction conditions with the polysaccharide yield as the evaluation index. Subsequently, CP was separated and purified through DEAE-52 cellulose column chromatography to obtain homogeneous polysaccharide fractions. On the basis of in vitro antioxidant activity screening, the structural properties were characterized via infrared spectroscopy, high-performance gel permeation chromatography, and ion chromatography, and animal experiments were performed to evaluate the anti-fatigue capacity of the optimal fraction. The results revealed that the optimal extraction parameters were 20 min of ultrasonication, a solid-to-liquid ratio of 1:20, and an ultrasonic power of 220 W, under which the maximum CP yield reached 8.55%. After purification, one neutral polysaccharide (CPN) and three acidic polysaccharides (CPA-1, CPA-2, CPA-3) were isolated. The yields of CPN, CPA-1, CPA-2, and CPA-3 were 40.3%, 22.6%, 13.7%, and 9.2% respectively, with their corresponding polysaccharide component recoveries being 78.3%, 89.6%, 65.4%, and 52.7%. The antioxidant activity of the four fractions followed the order: CPA-2>CPN>CPA-1>CPA-3. Given its high yield, favorable recovery efficiency and antioxidant activity comparable to CPA-2, CPN was selected for further investigation. Structural analysis confirmed that CPN possessed characteristic absorption peaks of polysaccharides, was mainly composed of glucose and mannose, and might contain two components with significantly different molecular weights. In vivo experiments demonstrated that CPN could significantly prolong the weight-bearing swimming time of mice, reduce the levels of blood urea nitrogen and blood lactic acid, and increase hepatic glycogen content, thereby verifying its anti-fatigue activity. These findings lay a solid foundation for the in-depth development and anti-fatigue application of Cistanche tubulosa polysaccharides.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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