羊栖菜多糖的提取和分离

羊栖菜多糖是从马尾藻科植物羊栖菜（Sargassum fusiforme(Harv.)Setchel)中提取到的，具有抗肿瘤，抗凝血，降血糖，提高机体免疫功能等生理活性，本文研究用水提法提取羊栖菜多糖，考虑到提取温度，时间，加水量等因素的影响，优化提取条件，确定水提法的最佳温度80℃，提取时间3h，水加量为羊栖菜样品的3倍，并采用乙醇沉淀，透析及凝胶层析等方法对羊栖菜提取物中多糖进行沉淀，提取和分离获得4个组分，并用光谱扫描法，凝胶色谱和旋光度法鉴定其纯度，进一步研究其分离获得的组分对生物体免疫功能指标的影响。     

Bioassay‐guided isolation of an active compound with protein tyrosine phosphatase 1B inhibitory activity from Sargassum fusiforme by high‐speed counter‐current chromatography

A rapid and efficient method using high‐speed counter‐current chromatography was established for the bioassay‐guided separation of an active compound with protein tyrosine phosphatase 1B inhibitory activity from Sargassum fusiforme. Under the bioassay guidance, the ethyl acetate extract with the best IC₅₀ value of 0.37 ± 0.07 μg/mL exhibited a potential protein tyrosine phosphatase 1B inhibitory activity, which was further separated by high‐speed counter‐current chromatography. The separation was performed with a two‐phase solvent system composed of n‐hexane/methanol/water (5:4:1, v/v). As a result, dibutyl phthalate (19.7 mg) with the purity of 95.3% was obtained from 200 mg of the ethyl acetate extract. Its IC₅₀ was 14.05 ± 0.06 μM, which was further explained by molecular docking. The result of molecular docking showed that dibutyl phthalate enfolded in the catalytic site of protein tyrosine phosphatase 1B. The main force between dibutyl phthalate and protein tyrosine phosphatase 1B was the hydrogen bond interaction with Gln266. In addition, hydrogen bond, van der Waals force and hydrophobic interaction with the amino acids (Ala217, Ile219, and Gly220) were also responsible for the stable protein‐ligand complex.     

Preparation of New Sargassum fusiforme Polysaccharide Long-Chain Alkyl Group Nanomicelles and Their Antiviral Properties against ALV-J

Avian leukosis virus subgroup J (ALV-J) is an immunosuppressive virus which has caused heavy losses to the poultry breeding industry. Currently, there is no effective medicine to treat this virus. In our previous experiments, the low-molecular-weight Sargassum fusiforme polysaccharide (SFP) was proven to possess antiviral activity against ALV-J, but its function was limited to the virus adsorption stage. In order to improve the antiviral activity of the SFP, in this study, three new SFP long-chain alkyl group nanomicelles (SFP-C12M, SFP-C14M and SFP-C16M) were prepared. The nanomicelles were characterized according to their physical and chemical properties. The nanomicelles were characterized by particle size, zeta potential, polydispersity index, critical micelle concentration and morphology. The results showed the particle sizes of the three nanomicelles were all approximately 200 nm and SFP-C14M and SFP-C16M were more stable than SFP-C12M. The newly prepared nanomicelles exhibited a better anti-ALV-J activity than the SFP, with SFP-C16M exhibiting the best antiviral effects in both the virus adsorption stage and the replication stage. The results of the giant unilamellar vesicle exposure experiment demonstrated that the new virucidal effect of the nanomicelles might be caused by damage to the phospholipid membrane of ALV-J. This study provides a potential idea for ALV-J prevention and development of other antiviral drugs.     

羊栖菜Sargassum fusiforme Setch繁殖与发育生物学的初步研究

对经济海藻羊栖菜的分类、形态结构、繁殖方式、发育方式、生态习性和人工育苗方法进行了探讨。     

高效毛细管电泳分析羊栖菜多糖对肿瘤机体红细胞合淌度的影响

为了分析羊栖菜多糖（SFPS）对肿瘤机体红细胞合淌度的影响,建立了肿瘤动物模型,分高、中、低剂量腹腔给予羊栖菜多糖7 d,采集并制备红细胞悬液,应用高效毛细管电泳法检测红细胞的合淌度。实验条件:毛细管为75 μm×50 cm,电泳缓冲液为含2 g/L羟丙基甲基纤维素的磷酸盐溶液(0.1 mol/L,pH 7.4),压力进样为3.448 kPa×10 s,分离电压为20 kV,柱温为25 ℃,电渗淌度为2.16×10-4 cm2·V-1·s-1。实验结果表明：羊栖菜多糖能降低肿瘤机体红细胞的电泳迁移时间(阴性对照组为15.76 min,SFPS低剂量组为13.96 min,SFPS中剂量组为12.90 min,SFPS高剂量组为13.51 min,正常对照组为11.51 min),增加红细胞的合淌度(阴性对照组为1.06×10-4 cm2·V-1·s-1,SFPS低剂量组为1.19×10-4 cm2·V-1·s-1,SFPS中剂量组为1.29×10-4 cm2·V-1·s-1,SFPS高剂量组为1.23×10-4 cm2·V-1·s-1,正常对照组为1.45×10-4 cm2·V-1·s-1),SFPS 3个剂量组红细胞的合淌度与阴性对照组比较均有非常显著的差异(P＜0.01)。羊栖菜多糖能够改变肿瘤机体红细胞的合淌度,并使之趋向于正常机体水平,这可能与其改变红细胞表面的电荷密度有关。高效毛细管电泳法可以作为检测红细胞生理状态和功能的一种辅助工具。     

微波消解-等离子体原子发射光谱法测定羊栖菜中的微量元素

采用HNO3-H2O2作消解剂对样品进行微波消解,利用等离子体原子发射光谱(ICP-AES)法同时测定羊栖菜中Fe、Mg、Ba、Ca、Mn和Al。对样品的前处理条件进行了探讨,通过加标回收实验,验证了分析数据的可靠性。在最佳测定条件下,方法检出限为0.0005~0.01μg/mL,相对标准偏差≤2.6%,加标回收率为95.7%~103.5%。该方法快速、简便、数据准确可靠,结果令人满意,适合常规分析。     

羊栖菜多糖对小鼠免疫功能的影响

采用从羊栖菜提取物分离获得的 4个组分中选择其中不同分子量 ,且得率较高的F1、f1组分 ,观察它们对小鼠免疫功能的影响 .结果显示按 4 0mg/(kg·d)剂量腹腔注射 7d后 ,对小鼠抗SRBC抗体生成有促进作用 ,同时明显提高小鼠脾指数 ,其中f1组最明显 .对f1组分按不同剂量注射 ,结果显示对小鼠胸腺T淋巴细胞增殖促进作用呈明显量效关系 .     

二维核磁共振对羊栖菜褐藻糖胶DJL的结构研究

羊栖菜是我国的特色海藻,褐藻糖胶是其主要生物活性成分. 葡萄糖醛酸甘露聚糖（DJL）是羊栖菜褐藻糖胶经脱硫和部分酸水解后的产物,本文采用2D NMR和甲基化分析等方法,对DJL的结构进行了研究. 结果显示,DJL主要由甘露糖（46.4%）和葡萄糖醛酸（50.5%）组成, 另外还含有少量的半乳糖,相对分子质量为11 KDa,其可能具有的一种结构为：β-D-GlcA-1→ [2)-α-D-Man(1→4)-β-D-GlcA(1→]_n1→4)-β-D-Gal(1→[4)-β-D-GlcA(1→2)-α-D-Man(1→]_n2→4)-β-D-GlcA(1→2)-α-D-Man.     

羊栖菜幼孢子体室内的培育

羊栖菜是浙江海区重要的大型栽培海藻 ,幼孢子体室内培育的结果将直接影响到栽培所需种苗的质量 .海区栽培的羊栖菜成熟期有一定的规律 ,温度、盐度是影响幼孢子体的生长和附着的重要因素 ,适宜的生长温度是在 2 0℃ ,附着的适宜温度是 15～ 2 0℃ ,适宜的盐度为 2 2‰～ 2 8‰ .     

羊栖菜“浙海1号”复水性与膨胀率的初步研究

比较研究了羊栖菜新品系“浙海1号”与本地传统栽培种的复水率与膨胀率的差异. 采用自然风干与不同温度下的干热鼓风干燥的方法. 获得“浙海1号”较适宜的干燥温度为50~60℃, “浙海1号”复水率、气囊直径、主枝直径分别高于本地传统栽培种21.7%、40.1%~45.9%、89.3%. 通过不同部位的复水率比较, 气囊是复水率最高的部分. “浙海1号”与本地传统栽培种相比, “浙海1号”膨胀率高出约为33%, 差异显著; 叶具有较高膨胀率且稳定性较强.