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元蘑多糖的结构特征及其调节巨噬细胞免疫活性机制

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[目的]为明确元蘑多糖的结构特征,并探究其调节巨噬细胞免疫活性机制.[方法]采用水提醇沉法获得元蘑多糖粗提物,并通过DEAE-52 离子交换层析及Sephacryl S-400 葡聚糖凝胶过滤层析对该多糖粗提物进行分离纯化,获得多糖组分SEP-0a.分别采用高效凝胶渗透色谱法、离子交换色谱法、傅立叶红外光谱法检测了SEP-0a的理化性质.进一步采用CCK-8 法、ELISA法、qPCR法、Western blot等方法考察了元蘑多糖SEP-0a对巨噬细胞RAW264.7 的免疫调节作用及机制.[结果]元蘑多糖SEP-0a由半乳糖、甘露糖、葡萄糖和岩藻糖组成,相对分子量为 4.23×104 Da,并且具有α构型与β构型.体外免疫活性结果表明,元蘑多糖SEP-0a可显著增强RAW264.7 细胞增殖和吞噬能力,促进该细胞活性氧(reactive oxygen species,ROS)、一氧化氮(nitric oxide,NO)的分泌,提高肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6 的表达,Western blot检测发现,该多糖可显著提高核转录因子-κB(nuclear factor-κB,NF-κB)信号通路p65 蛋白磷酸化表达.[结论]元蘑多糖SEP-0a可通过NF-κB信号通路调控巨噬细胞的免疫活性.
Structural Characteristics of Sarcomyxa edulis Polysaccharide and Its Mechanism of Regulating Macrophage Immunomodulatory Activity
[Objective]In order to clarify the structural characteristics of Sarcomyxa edulis polysaccharides and explore its mechanism of regulating macrophage immunomodulatory activity.[Method]The crude polysaccharide of S.edulis was obtained by water extraction and ethanol precipitation,and the purified polysaccharide component SEP-0a was obtained by DEAE-52 ion exchange column chromatography and Sephacryl S-400 glucan gel column chromatography.The physicochemical properties of SEP-0a were investigated by high performance gel permeation chromatography evaporative light-scattering detector,ion exchange chromatography high performance liquid chromatography and Fourier transform infrared spectrometer.On this basis,the immunomodulatory effect of polysaccharide SEP-0a from S.edulis on macrophage cell RAW264.7 was further studied by CCK-8,ELISA,qPCR,Western Blot and other methods.[Result]The S.edulis polysaccharide SEP-0a was composed of galactose,mannose,glucose and fucose with a relative molecular weight of 4.23×104 Da,and had α and β configurations.The results of in vitro immunocompetence showed that S.edulis polysaccharide SEP-0a significantly increased the proliferation and phagocytic ability of RAW264.7 cells,promoted the secretion of reactive oxygen species(ROS),nitric oxide(NO)and increased the expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6 in these cells.Western blot results showed that this polysaccharide significantly increased p65 protein phosphorylation expression in the nuclear factor-κB(NF-κB)signaling pathway.[Conclusion]The polysaccharide SEP-0a of S.edulis may regulates the immune activity of macrophages by NF-κB signal pathway.

Sarcomyxa edulis polysaccharidestructural characteristicsRAW264.7 cellsNF-κB signal pathway

王子璇、隋玉、尚学钰、马思嘉、吴天香、刘洋、王琦

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吉林农业大学食药用菌教育部工程研究中心,长春 130118

吉林农业大学植物保护学院,长春 130118

吉林农业大学中药材学院,长春 130118

元蘑多糖 结构特征 RAW264.7 细胞 NF-κB信号通路

中俄菌物资源评价与利用国家食用菌产业技术体系

20220402051GCARS-20-08B

2024

生物技术通报
中国农业科学院农业信息研究所

生物技术通报

CSTPCD北大核心
影响因子:0.505
ISSN:1002-5464
年,卷(期):2024.40(1)
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