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目的:对鹿茸糖胺聚糖不同提取方法的效果进行比较,并对其免疫调节和抗氧化活性进行研究。方法:分别采用蛋白酶水解法、稀碱-蛋白酶水解法提取鹿茸糖胺聚糖,随后选用三氯乙酸法或Sevage法除蛋白。以提取物的基本理化性质、总糖胺聚糖、己糖醛酸和蛋白质含量为指标对4种提取方法进行比较。应用WST-8法检测鹿茸糖胺聚糖对小鼠脾淋巴细胞增殖的影响,以Fe3+还原能力、DPPH自由基及羟自由基清除能力评价鹿茸糖胺聚糖的抗氧化活性。结果:采用胰蛋白酶-木瓜蛋白酶水解,三氯乙酸法除蛋白,制备所得鹿茸糖胺聚糖纯度最高。鹿茸糖胺聚糖对ConA诱导的小鼠脾T淋巴细胞增殖呈倒U型曲线;对LPS诱导的小鼠脾B淋巴细胞增殖有抑制作用。鹿茸糖胺聚糖具有Fe3+还原能力、DPPH自由基及羟自由基清除能力。结论:蛋白酶解法适用于鹿茸糖胺聚糖的提取,鹿茸糖胺聚糖可能具有双向免疫调节作用,并具有良好的抗氧化作用。
OBJECTIVE: To compare the effects of different extraction methods of deer antler glycosaminoglycan, and to study its immunoregulation and antioxidant activity. Methods: The antler glycosaminoglycan was extracted by protease hydrolysis and dilute alkali - protease hydrolysis respectively, then the trichloroacetic acid method or Sevage method was used to remove protein. The four extraction methods were compared with the basic physical and chemical properties, total glycosaminoglycan, hexuronic acid and protein content of the extract as an index. WST-8 method was used to detect the antler glycosaminoglycan on mouse splenic lymphocyte proliferation, Fe3 + reducing ability, DPPH free radical and hydroxyl radical scavenging ability to evaluate the antler glycosaminoglycan activity. Results: Trypsin-papain hydrolysis, trichloroacetic acid method of protein removal, the highest purity of the resulting deer antler glycosaminoglycan. Antler glycosaminoglycan on ConA-induced mouse splenic T lymphocyte proliferation was inverted U-shaped curve; LPS-induced mouse spleen B lymphocyte proliferation was inhibited. Deer velvet glycosaminoglycan with Fe3 + reducing power, DPPH free radicals and hydroxyl radical scavenging capacity. Conclusion: The proteolytic method is suitable for the extraction of deer antler glycosaminoglycan. The antler glycosaminoglycan may have two-way immunomodulatory effect and good anti-oxidant effect.