College of Food and Pharmacy, Zhejiang Ocean University, 1 South Haida Road, Zhoushan, 316000, People's Republic of China.
College of Food and Pharmacy, Zhejiang Ocean University, 1 South Haida Road, Zhoushan, 316000, People's Republic of China; Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, 1 South Haida Road, Zhoushan, 316000, People's Republic of China.
J Ethnopharmacol. 2020 Jul 15;257:112833. doi: 10.1016/j.jep.2020.112833. Epub 2020 Apr 11.
Shiraia bambusicola is a parasitic fungus on the twigs of bamboos. Its relatively large stroma has high medicinal value and can treat a variety of diseases such as rheumatoid arthritis, cold stomach pain, sciatica, injuries, chronic bronchitis, and infantile. It is widely distributed in many provinces in Southern China and also is also found in Japan.
Medicinal fungi were important resources for bioactive polysaccharides. To explore bioactive polysaccharides from Shiraia bambusicola, a heteropolysaccharide SB2-1 was purified and obtained from S. bambusicola and its immunostimulating activity was researched.
The polysaccharide from S. bambusicola was extracted and purified using enzyme assisted extraction, ethanol precipitation, anion-exchange and size-exclusion chromatography. Molecular weight of polysaccharide was estimated by high performance gel permeation chromatography. Monosaccharide compositions were determined by high performance liquid chromatography after pre-column derivatization and UV detection. Structure information was elucidated by IR spectrum, GC-MS analysis after methylation and gradual acid hydrolysis of the polysaccharide. The RAW264.7 cells were used to study the immunostimulating activity in vitro.
Physicochemical and structural analyses showed that SB2-1 was a neutral heteropolysaccharide with molecular weight at 22.2 kDa and consisted of glucose, galactose and mannose at a ratio of 2.0:1.5:1.0. The structure of SB2-1 was a branched polysaccharides composed of a mannan core and side chains consisted of glucose and galactose. The mannan core was composed of (1→2)-Manp as the main chain. Glucose with (1→4)-D-Glcp, (1→2)-D-Glcp and (1→6)-D-Glcp at different degrees of polymerization were linked at C-6 and C-3 of the (1→2)-Manp as the side chains. The galactose with the linages of (1→6)-D-Galf, →2)-D-Galf(1→ and terminal D-Galf(1→ also existed in the side chain. The study on the immunostimulating activities of SB2-1 and its core structure P-2 were investigated on RAW264.7 macrophages. The results showed that SB2-1 could activate RAW264.7 macrophage and significantly improve its phagocytic ability by neutral red uptake experiment. Meanwhile, SB2-1 increased significantly higher inducible nitric oxide synthase (iNOS) production and the productions of IL-1, IL-6, IL-12 and TNF-α. The effect of SB2-1 was better than its core structure P-2 produced by gradual acid hydrolysis, which meant the side chains played an important role in the immunostimulating activities.
The investigation demonstrated that the galactofuranose-containing mannogalactoglucan was characteristic polysaccharides in S. bambusicola and could enhance the activation of macrophages.
密环菌是一种寄生在竹枝上的真菌。其相对较大的菌核具有很高的药用价值,可治疗类风湿性关节炎、胃寒腹痛、坐骨神经痛、跌打损伤、慢性支气管炎、婴儿腹泻等多种疾病。它广泛分布于中国南方的许多省份,也在日本发现。
药用真菌是生物活性多糖的重要资源。为了从密环菌中探索生物活性多糖,从密环菌中提取并纯化了一种杂多糖 SB2-1,并研究了其免疫刺激活性。
采用酶辅助提取、乙醇沉淀、阴离子交换和凝胶渗透色谱法从密环菌中提取和纯化多糖。采用高效凝胶渗透色谱法估计多糖的分子量。多糖的单糖组成采用柱前衍生化和紫外检测后的高效液相色谱法测定。采用红外光谱、GC-MS 分析和多糖逐步酸水解后甲基化对其结构信息进行阐明。采用 RAW264.7 细胞体外研究其免疫刺激活性。
理化分析和结构分析表明,SB2-1 是一种中性杂多糖,分子量为 22.2 kDa,由葡萄糖、半乳糖和甘露糖组成,比例为 2.0:1.5:1.0。SB2-1 的结构是一种支链多糖,由甘露聚糖核心和由葡萄糖和半乳糖组成的侧链组成。甘露聚糖核心由(1→2)-Manp 作为主链。葡萄糖以不同程度的聚合度(1→4)-D-Glcp、(1→2)-D-Glcp 和(1→6)-D-Glcp 连接在 C-6 和 C-3 的(1→2)-Manp 上作为侧链。半乳糖侧链中也存在(1→6)-D-Galf、→2)-D-Galf(1→和末端 D-Galf(1→的连接。通过中性红摄取实验研究了 SB2-1 及其核心结构 P-2 对 RAW264.7 巨噬细胞的免疫刺激活性。结果表明,SB2-1 能激活 RAW264.7 巨噬细胞,显著提高其吞噬能力。同时,SB2-1 显著增加诱导型一氧化氮合酶(iNOS)的产生以及 IL-1、IL-6、IL-12 和 TNF-α的产生。SB2-1 的作用优于其通过逐步酸水解产生的核心结构 P-2,这意味着侧链在免疫刺激活性中起着重要作用。
研究表明,含有半乳糖呋喃糖的甘露半乳糖葡聚糖是密环菌中特征性多糖,能增强巨噬细胞的激活。