Macrophage-stimulating activities of newly isolated complex polysaccharides from Parachlorella kessleri strain KNK-A001
Macrophage-stimulating activities of newly isolated complex polysaccharides from Parachlorella kessleri strain KNK-A001
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从凯斯氏副小球藻菌株 KNK-A001 中新分离出的复合多糖的巨噬细胞刺激活性
DOI：
10.1016/j.ijbiomac.2017.06.014
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发表时间：
2017
影响因子：
8.2
通讯作者：
Oda Tatsuya
中科院分区：
文献类型：
--
作者：
Ueno Mikinori;Cho Kichul;Hirata Narumi;Yamashita Kenji;Yamaguchi Kenichi;Kim Daekyung;Oda Tatsuya
本课题组前期的研究表明，克氏副小球藻（KNK-A001）具有免疫刺激活性，腹腔注射后可提高小鼠自然杀伤（NK）细胞活性，口服给药后对病毒感染模型虾具有保护作用。在本研究中，我们试图了解KNK-A001的组成物质，这些物质负责免疫刺激活性。首先，我们通过DEAE阴离子交换层析从KNK-A001中分离得到五个多糖组分。其中，F5表现出最有效的诱导一氧化氮（NO）的分泌在RAW 264.7细胞中，诱导型NO合酶（iNOS）的mRNA和蛋白质表达水平在F5处理的RAW 264.7细胞增加。在F5处理的RAW 264. 7细胞中观察到核因子-κ B（NF-κB）的p65亚基的核转位显著增加。F5还可诱导RAW264.7细胞分泌肿瘤坏死因子（TNF）-α。用丝裂原活化蛋白激酶（MAP激酶）抑制剂分析表明，c-Jun N-末端激酶（JNK）和p38 MAP激酶主要参与F5诱导的NO和TNF-α的产生。F5的成分分析鉴定出主要成分为半乳糖、葡萄糖、半乳糖醛酸和甘露糖。凝胶过滤分析表明F5的分子量约为400 kDa。
Our previous studies demonstrated that the microalgaParachlorella kessleri(KNK-A001) has immunostimulatory activities, which were observed as an increase in natural killer (NK) cell activity in mice after intraperitoneal injection or as a protective effect on a virus-infected model shrimp after oral administration. In this study, we attempted to gain insight into the constituent substances of KNK-A001 that are responsible for the immunostimulatory activity. First, we obtained five polysaccharide fractions from KNK-A001 by DEAE anion exchange chromatography. Among the fractions, F5 showed the most potent induction of nitric oxide (NO) secretion in RAW264.7 cells, and both mRNA and protein expression levels of inducible NO synthase (iNOS) were increased in F5-treated RAW264.7 cells. A significant increase in the nuclear translocation of the p65 subunit of nuclear factor-kappa B (NF-κB) was observed in F5-treated RAW264.7 cells. F5 also induced the secretion of tumor necrosis factor (TNF)-α in RAW264.7 cells. Analysis using mitogen-activated protein (MAP) kinase inhibitors suggested that c-Jun N-terminal kinase (JNK) and p38 MAP kinase were mainly involved in F5-induced NO and TNF-α productions. The compositional analysis of F5 identified the main constituents as galactose, glucose, galacturonic acid, and mannose. Gel-filtration analysis suggested that molecular mass of F5 was approximately 400 kDa.