仿刺参卵和体壁多肽的制备及免疫活性

Preparation of Peptides from Eggs and Body Wall of Apostichopus japonicus and Their Immunoenhancing Effect

以仿刺参卵和体壁为原料制备多肽,测定其基本营养成分和分子质量分布,并研究其对淋巴细胞增殖作用和小鼠免疫功能的影响。采用生物酶解技术和切向流超滤技术分离和制备仿刺参卵和体壁多肽,磺酰罗丹明B比色分析法测定了多肽质量浓度(10、50、100、500μg/mL)对小鼠淋巴细胞增殖(spleen lymphocyte proliferation,SLP)能力的影响,设定水对照组和多肽低、中、高(日剂量83.3、166.7、500.0 mg/kg)剂量组进行30 d小鼠灌胃实验,检测了小鼠脏器/体质量、迟发型变态反应能力、伴刀豆球蛋白A诱导的小鼠脾淋巴细胞转化能力、抗体生成细胞数、小鼠血清溶血素水平、小鼠碳廓清能力、小鼠腹腔巨噬细胞吞噬鸡红细胞能力及自然杀伤细胞活力等指标。结果表明,仿刺参卵和体壁1~10 kDa多肽(EP1和BWP1)在500μg/mL时均对SLP具有显著的促进作用,其粗蛋白含量分别为64.74、70.25 g/100 g,氨基酸总量分别为45.69、63.26 g/100 g,分子质量分别分布在130~1 600 Da及130~2 500 Da之间。经口服灌胃给予小鼠不同剂量的多肽30 d,EP1可提高小鼠的细胞免疫功能和单核-巨噬细胞吞噬功能,BWP1可提高小鼠的体液免疫功能和单核-巨噬细胞吞噬功能。结果提示EP1和BWP1均具有增强免疫力的功能,可开发为新的免疫调节产品。

The nutritional components and molecular mass distribution of the peptides prepared from the eggs and body wall of Apostichopus japonicus were detected, and then their effects on spleen lymphopoiesis and immune function in mice were evaluated. Four peptides from the eggs and body wall of sea cucumber were prepared by enzymatic hydrolysis and crossflow ultrafiltration. Mouse spleen lymphocyte proliferation （SLP） in vitro in the presence of each of the peptides （10, 50, 100 and 500 μg/mL） was measured by the sulforhodamine B colorimetric method. Furthermore, mice were subjected to oral gavage with each of the four peptides at low, middle and high dosages （83.3, 166.7 and 500.0 mg/kg） for 30 consecutive days. Those given water were used as a control group. After the experiments, organ/body mass ratio, delayed-type hypersensitivity, T lymphocyte proliferation induced by concanavalin A, the number of antibodyforming cells, serum hemolysin formation, carbon clearance capability, chicken red blood cell phagocytosis by mouse macrophages and natural killer cell activity were measured. The results showed that EP1 and BWP1 （1–10 kDa） at 500 μg/mL had a significant effect on SLP. The crude protein contents of EP1 and BWP1 were 64.74 and 70.25 g/100 g, the total amino acids were 45.69 and 63.26 g/100 g, and the molecular mass distribution was 130–1 600 and 130–2 500 Da, respectively. After the mice were orally administered with different doses of the peptides for 30 days, both EP1 and BWP1 enhanced monocyte-derived macrophage phagocytosis; EP1 improved cellular immune function while BWP1 augmented humoral immune function. In summary, EP1 and BWP1 from the eggs and body wall from Apostichopus japonicus had immunoenhancing effects and could be developed as new immunomodulatory products.