阿魏侧耳胞外多糖分离纯化及其免疫活性研究  

Purification and immunomodulation activity of exopolysaccharides from Pleurotus ferulae Lanzi

作　　者：王晶[1,2,3] 乔洁 裴新云[1] 常世民 刘学娟 闫训友[1,2,3] WANG Jing;QIAO Jie;PEI Xinyun;CHANG Shimin;LIU Xuejuan;YAN Xunyou(College of Life Sciences,Langfang Normal University,Langfang 065000,China;Technical Innovation Center for Utilization of Edible and Medicinal Fungi in Hebei Province,Langfang 065000,China;Langfang Key Laboratory of Cell Engineering and Application,Langfang 065000,China)

机构地区：[1]廊坊师范学院生命科学学院,河北廊坊065000 [2]河北省食药用菌资源高值利用技术创新中心,河北廊坊065000 [3]廊坊市细胞工程与应用研究重点实验室,河北廊坊065000

出　　处：《食品与发酵工业》2021年第18期127-134,共8页Food and Fermentation Industries

基　　金：河北省自然科学基金青年基金项目(C20184080059);河北省高等学校科学技术研究重点项目(ZD2019068);河北省高等教育教学改革研究与实践项目(2020GJJG266);河北省大学生创新创业训练计划项目(S202010100019)。

摘　　要：研究了阿魏侧耳(Pleurotus ferulae Lanzi)胞外多糖的分离纯化及其对巨噬细胞和脾淋巴细胞的免疫调节作用。采用醇沉、透析、冷冻干燥、DEAE-52柱层析和Sephadex G-100柱层析等方法从发酵液分离纯化阿魏侧耳胞外多糖,通过高效凝胶渗透色谱法与高效液相色谱法测定其分子质量和单糖组成,以灌洗腹腔法和密度梯度离心法分别收集巨噬细胞和脾淋巴细胞,并利用荧光探针法和酶联免疫吸附实验分别测定体外培养的巨噬细胞中NO和活性氧含量及2种细胞培养上清中多种免疫相关细胞因子的浓度。结果表明,阿魏侧耳胞外多糖水提组分PFEPw-1的分子质量为64.2 kDa,由D-甘露糖、D-葡萄糖、D-半乳糖和L-阿拉伯糖组成,含量比为2.42∶3.90∶2.27∶1.41。终浓度为10~30 mg/L的PFEPw-1可显著提高巨噬细胞对NO和活性氧的合成(P<0.05)及肿瘤坏死因子-α和干扰素-γ的分泌量(P<0.05),20~30 mg/L的PFEPw-1可显著提高巨噬细胞对白细胞介素-1和白细胞介素-6的分泌量(P<0.05)及脾淋巴细胞对白细胞介素-2和干扰素-γ的分泌量(P<0.05)。该研究可为阿魏侧耳胞外多糖的工业化生产及开发应用提供理论支持。To investigate the optimum fermentation medium of exopolysaccharides from Pleurotus ferulae Lanzi,isolation and purification of water extract component in addition to its immunomodulation activity on macrophages and spleen lymphocytes were cultured in vitro.And ethanol precipitation,dialysis,lyophilization,DEAE-cellulose 52 chromatography and Sephadex G-100 chromatography were carried out to obtain water extract component of exopolysaccharides from P.ferulae Lanzi(PFEPw-1).The molecular weight and monosaccharide composition of PFEPw-1 were determined by high-performance gel permeation chromatography and high-performance liquid chromatography,respectively.Macrophages and spleen lymphocytes were isolated using the peritoneal lavage method and density gradient centrifugation,respectively.Fluorescent probe technique and enzyme-linked immunosorbent assay were carried out to measure the content of intracellular nitric oxide and reactive oxygen,and the levels of immune-related cytokines in culture supernatant,respectively.The results showed that PFEPw-1 was composed of D-mannose,D-glucose,D-galactose and L-arabinose L-fucose with a content ratio of 2.42∶3.90∶2.27∶1.41,and the molecular weight of PFEPw-1 was 64.2 kDa.Compared with the control group,10-30 mg/L PFEPw-1 enhanced intracellular NO and reactive oxygen content(P<0.05),and improved the secretion level of tumor necrosis factor-αand interferon-γin the culture supernatant(P<0.05)of macrophages cultured in vitro.Meanwhile,20-30 mg/L PFEPw-1 could improve the secretion level of interleukin-1 and interleukin-6 in the culture supernatant of macrophages(P<0.05),and interleukin-2 and interferon-γin the culture supernatant of spleen lymphocytes(P<0.05).The research provides theoretical support for industrial production,development and utilization of exopolysaccharides from P.ferulae Lanzi.

关 键 词：阿魏侧耳 胞外多糖 巨噬细胞 淋巴细胞 免疫调节 

分 类 号：R285[医药卫生—中药学]