鹿茸糖胺聚糖提取方法的比较及其活性研究

Extraction methods and activity of glycosaminoglycan from velvet antler

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 on the extraction of glycosaminoglycan from velvet antler, and study the immunomodulatory and antioxidant activity of glycosaminoglycan. Methods： Glycosaminoglycan was extracted from velvet antler by protease hydrolysis method and dilute alkali-protease hydrolysis method, respectively. Trichloroacetic acid method and Sevage method were used to remove protein from the crude glycosaminoglycan. Basic physical and chemical properties, total glycosaminoglycan, hexuronic acid and protein content were used to evaluate the effect of four extraction methods. The effect of glycosaminoglycan on the proliferation of murine splenocytes was detected by WST-8. Fe3~ reduction ability, DPPH radical and hydroxyl radical scavenging capacity were used to evaluate the antioxidant activity of glycosaminoglycan from velvet antler. Results： Glycosaminoglycan that extracted by trypsin-papain hydrolysis, following deproteinization with trichloroacetic acid, had the highest purity. The relationship between the concentration of glycosaminoglycan and effect on ConA-stimulated splenocyte proliferation showed an inverted U-shaped curve. Glycosaminoglycan inhibited the proliferation of LPS-stimulated murine splenocytes. Glycosaminoglycan had Fe3~ reduction ability, DPPH radical and hydroxyl radical scavenging capacity. Conclusions： Protease hydrolysis method is applicable to extract glycosaminoglycan from velvet antler. Glycosaminoglycan from velvet antler has bidirectional immunomodulatory effect and antioxidant activity.