绣球菌多糖及其体外免疫活性研究

张迪; 王宏雨; 肖冬来; 林衍铨

doi:10.19303/j.issn.1008-0384.2019.09.015

绣球菌多糖及其体外免疫活性研究

doi: 10.19303/j.issn.1008-0384.2019.09.015

福建省农业科学院食用菌研究所，福建　福州　350014

基金项目: 福建省科技计划公益类专项（2018R1020-3）；福建省财政专项——福建省农业科学院科技创新团队建设项目（STIT2017-1-6）

详细信息

作者简介:
张迪（1983−），男，硕士，副研究员，主要从事食用菌天然产物与加工技术研究

通讯作者:
林衍铨（1963−），男，研究员，主要从事食（药）用菌栽培研究（E-mail：lyq-406@163.com）

中图分类号: S 646.9
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出版历程
- 收稿日期: 2019-04-19
- 修回日期: 2019-09-09
- 刊出日期: 2019-09-01

Chemistry and In Vitro Immunological Activity of Polysaccharides from Sparassis latifolia

Institute of Edible & Medicinal Fungi, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350014, China

摘要

摘要: 目的分析绣球菌水溶性多糖的结构与免疫活性。方法采用热水提取法从广叶绣球菌Sparassis latifolia的真空冷冻干燥品中提取得到绣球菌水溶性多糖SCG-D，然后通过DEAE Sepharose Fast Flow离子交换层析对其进行分离，获得SCG-N和SCG-A两个多糖组分，并对SCG-A组分进行HPSEC、单糖组成分析、红外光谱、¹H-NMR和¹³C-NMR核磁共振的结构分析。通过大鼠脾淋巴细胞体外刺激活性测试比较了SCG-D及其组分SCG-A、SCG-N的体外免疫活性差异。结果绣球菌水溶性多糖SCG-D具有促进鼠脾淋巴细胞增殖的活性，其酸性多糖组分SCG-A促进作用最强，其100 μg·mL⁻¹ 72 h处理组的增殖率为32.7%，而SCG-N多糖组分的活性较SCG-A低，其100 μg·mL⁻¹ 72 h处理组增殖率为11.66%。结构分析结果表明SCG-A的重均分子量为4.30×10⁵ Da，其主链结构为α-1,4-D-葡聚糖，主要由葡萄糖和半乳糖构成，并且还含有一定量的1-6分支结构。结论绣球菌冻干品的水溶性多糖及其组分具有促进大鼠脾淋巴细胞体外增殖的免疫活性，其中活性最强的酸性多糖组分SCG-A是一种具有1-6分支结构的α-1,4-葡聚糖。
- 广叶绣球菌 /
- 多糖 /
- 脾淋巴细胞 /
- 免疫
Abstract: Objective Chemical structure and in vitro immunological activity of the water soluble polysaccharides from Sparassis latifolia were studied . Method The lyophilized powder of S. latifolia fruiting bodies was extracted by hot water to obtain polysaccharides SCG-D. From the extract, the SCG-N and SCG-A fractions were isolated by the DEAE-Sepharose Fast Flow ion exchange chromatography. The structure of SCG-A was analyzed using HPSEC, monosaccharide composition analysis, FT-IR, and NMR. The in vitro immunological activities of the extract and the fractions were determined by a test on rat spleen lymphocytes. Result SCG-D was found active in promoting the lymphocyte proliferation. SCG-A exhibited the strongest activity with a proliferation rate of 32.7% at the concentration of 100 μg/mL in 72h, while SCG-N activity was lower at 11.66%. The molecular weight of SCG-A was 4.30×10⁵Da with an α-1,4-D-glucan as its main chain consisting primarily of glucose and galactose with some 1-6-branches. Conclusion The water-soluble polysaccharides and its fractions obtained from the lyophilized S. latifolia powder showed varying in vitro immune activities of promoting the proliferation of rat spleen lymphocytes. The acidic SCG-A had a main chain structure of α-1,4-glucan with 1-6-branches and showed the greatest immunological activity among the 3 polysaccharide materials.
- Sparassis latifolia /
- polysaccharide /
- spleen lymphocyte /
- immunity

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图 1 绣球菌SCG-D多糖DEAE Sepharose Fast Flow的洗脱曲线

Figure 1. Elution curve of SCG-D on DEAE Sepharose Fast Flow resin

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图 2 SCG-A和SCG-N的HPSEC凝胶色谱图

Figure 2. HPSEC gel chromatography of SCG-A and SCG-N fractions

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图 3 标样与SCG-A水解样品PMP衍生物的色谱图

注：1：甘露糖；2：鼠李糖；3：葡萄糖醛酸；4：半乳糖醛酸；5：葡萄糖；6：半乳糖；7：木糖。

Figure 3. Chromatography of PMP derivatives of monosaccharide standards and SCG-A hydrolysate

Note: 1: Man; 2: Rha; 3: GlcU; 4: GalU; 5: Glu; 6: Gal; and, 7: Xyl.

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图 4 SCG-A的红外吸收光谱图

Figure 4. FT-IR spectrum of SCG-A

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图 5 SCG-A的¹H-NMR谱图

Figure 5. ¹H-NMR spectrum of SCG-A

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图 6 SCG-A的¹³C-NMR谱图

Figure 6. ¹³C-NMR spectrum of SCG-A

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图 7 绣球菌多糖SCG-A对大鼠脾淋巴细胞增殖的影响

注：** P＜0.01，图8-9同。

Figure 7. Effect of SCG-A on in vitro proliferation of rat spleen lymphocytes (** P < 0.01)

Note: ** P＜0.01，the same as Fig. 8-9.

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图 8 绣球菌多糖SCG-N对大鼠脾淋巴细胞增殖的影响

Figure 8. Effect of SCG-N on in vitro proliferation of rat spleen lymphocytes

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图 9 绣球菌多糖SCG-D对大鼠脾淋巴细胞增殖的影响

Figure 9. Effect of SCG-D on in vitro proliferation of rat spleen lymphocytes

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图 10 不同绣球菌多糖处理对大鼠脾淋巴细胞的增殖率的影响（72 h处理，MTT法）

Figure 10. Effects of polysaccharide extract and fractions on in vitro proliferation rate of rat spleen lymphocytes (72 h, by MTT)

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