海带岩藻聚糖及其级分对体外培养肉鸡巨噬细胞免疫功能的影响

Effects of Fucoidan and Its Fraction Isolated from Laminaria japonica on Immunity of Broiler Macrophages in vitro

【目的】通过研究海带岩藻聚糖及其级分对肉鸡巨噬细胞免疫功能的影响,寻找海带提取物调节肉鸡免疫功能的有效级分,为饲料工业科学使用海带资源提供理论依据。【方法】采用水提醇沉的方法,从海带中提取岩藻聚糖,通过纤维柱和凝胶柱层析,对岩藻聚糖进行分级纯化。体外培养肉鸡腹腔巨噬细胞,在培养液中分别加入岩藻聚糖(200μg·ml-1)和岩藻聚糖5种级分(100μg·ml-1),采用MTT方法检测细胞存活率,采用ELISA检测细胞因子IL-1和TNF-α,同时测定巨噬细胞的吞噬活性等其它免疫指标。【结果】海带岩藻聚糖及其级分显著提高肉鸡腹腔巨噬细胞的吞噬活性,促进巨噬细胞呼吸爆发、NO的产生量和iNOS活性,促进巨噬细胞分泌IL-1(P<0.05);海带岩藻聚糖不同级分对肉鸡巨噬细胞免疫功能的调控作用不同,其中级分Ⅲ和级分Ⅳ的促进作用较强,与对照组相比,可显著提高巨噬细胞的吞噬活性、呼吸爆发、NO产生量和iNOS活性。对其它指标无显著影响(P>0.05)。【结论】海带岩藻聚糖及其级分可提高体外培养肉鸡腹腔巨噬细胞的免疫功能,不同级分对肉鸡巨噬细胞免疫功能的调控作用不同,其中级分Ⅲ和级分Ⅳ促进巨噬细胞免疫功能的作用最强,是海带岩藻聚糖中的有效级分。

[ Objective ] The immunomodulatory activities of a Fucoidan and its fraction, purified from the Laminaria japonica （Fucoidan from Laminaria japonica, FLJ）, were investigated in vitro in relation to broiler peritoneal macrophages. [Method] Fucoidan was extracted from Laminaria japonica and used ion exchange and size exclusion chromatography to separate them into five fractions. Analysis of the effects of Fucoidan and five fractions on broilers peritoneal macrophages immunity function was conducted. The broiler macrophages were cultured with fucoidan （200 μg·ml^-1） and fraction from I to V （100 μg·ml^-1） for 24 h, the cell vialibility was detected by using the mothod of MTT and the phagocytic activity was detected. The induced nitric oxide synthase （iNOS）, NO, IL-1 and TNF-α in culture supernatants was measured. The respiratory burst capacity in macrophages was measured by luminal dependent chemiluminescence. [Result] Cell vialibility results showed that the proliferation of macrophages was not influenced by FLJ and its fractions. The treatment of FLJ and its fraction increased the phagocytic activity, enhanced the production of IL-1, NO, iNOS and the respiratory burst capacity （P〈0.05）. The fucoidan and its fraction had no significant effect on TNF-α（P 〉0.05）. The different fractions had different effects on broiler macrophage immunity function. Fraction Ⅲ and fraction Ⅳ had the most effective promoting action. Their promotion effect was higher than fraction V. The fraction V had no effect on broilers macrophages immunity function. [Conclusion] The fucoidan isolated from Laminaria japonica had promoting effect on broiler macrophage immunity at 200 μg·ml^-1 The fraction Ⅲ and fraction Ⅳ purified from fucoidan were the effective ftaction among the five fractions.