红毛藻多糖的化学组成及其体外免疫诱导活性研究
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福建省自然科学基金项目(2015J01140);国家自然科学基金项目(31501441)


Studies on Composition, in Vitro Immune-induced Activities of Polysaccharides Isolated from Bangia fusco-purpure
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    摘要:

    目的:对红毛藻中多糖组分进行提取、分离纯化,分析其化学组成和体外免疫诱导活性,阐明红毛藻具有提高免疫力食药用功效机制,为红毛藻的高值化利用提供科学依据。方法:通过水提醇沉法从红毛藻中提取红毛藻粗多糖,经阳离子交换柱层析和Sephadex G75柱层析纯化得到红毛藻多糖(BFP),再用DEAE-cellulose 52柱层析对BFP进行分级纯化得到3个多糖组分F1、F2和F3;采用PMP柱前衍生、HPLC及化学方法分析其单糖组成和化学成分;通过RAW264.7细胞模型分析BFP、F1、F2和F3的体外免疫诱导活性和其相关细胞信号传导通路。结果:组成分析结果表明BFP、F1、F2和F3均为杂多糖且单糖组成存在较大差异。细胞模型结果表明,BFP、F1、F2和F3在所测定质量浓度范围(0~100 μg/mL)均显著诱导RAW264.7细胞活化,释放NO和分泌TNF-α,而不诱导ROS的产生。细胞信号传导通路抑制试验结果表明BFP具有体外免疫诱导活性与细胞的NF-κB、JNK MAPK、ERK MAPK和p38 MAPK信号通路的激活相关;F1与细胞的NF-κB、JNK MAPK、ERK MAPK和p38 MAPK信号通路的激活有关;而F2和F3作用机制相似,分别与细胞的NF-κB、JNK MAPK和ERK MAPK信号通路的激活相关。结论:红毛藻多糖具有显著的体外免疫诱导活性,其中分级纯化多糖组分F1和F2是BFP中主要的免疫诱导活性组分;BFP、F1、F2和F3诱导RAW264.7细胞活化释放NO的共同的信号传导通路主要有两条:即NF-κB和JNK MAPK信号途径;而诱导RAW264.7细胞分泌TNF-α的共同信号通路主要是JNK MAPK和ERK MAPK信号途径。

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    Objective: In this study, the polysaccharide derived from Bangia fusco-purpurea was separated and purified, and its chemical composition and in vitro immune-induced activity were analyzed to illuminate food and medicine mechanism of Bangia fusco-purpurea, which provided scientific basis for the high-efficient utilization of Bangia fusco-purpurea. Methods: Crude polysaccharide was extracted from Bangia fusco-purpurea by hot water extraction and ethanol precipitation. The polysaccharide fractions (F1, F2 and F3) were obtained with isolating and purifying polysaccharide BFP by cation exchange column chromatography, Sephadex G75 chromatography and DEAE-cellulose 52 exchange chromatography. The monosaccharide composition and chemical component of BFP, F1, F2 and F3 were analyzed by PMP pre-column derivatization high performance liquid chromatography (HPLC) and the chemical methods. Further, the in vitro immune-induced activity and signal transduction pathway of BFP, F1, F2 and F3 were analyzed by mouse macrophage RAW264.7 cells model. Results: The results of chemical component and monosaccharide composition showed that BFP, F1, F2 and F3 were heteropolysaccharides with significant differences in monosaccharide compositions. The in vitro immune-induced activity cells model revealed that BFP, F1, F2 and F3 significantly induced RAW264.7 cells activation to release NO and TNF-α in the measured concentration range (0~100 μg/mL), but did not induce the generation of ROS. Cellular signal pathway inhibition experimental results show that the immune-induced activity of BFP was related to the cellular signal transduction pathways activation of NF-κB, JNK MAPK, ERK MAPK, and p38 MAPK; F1 was associated with the cellular signal transduction pathways activation of NF-κB, JNK MAPK, ERK MAPK and p38 MAPK; however, the mechanisms of action of F2 and F3 were similar, which were related to the activation of NF-κB, JNK MAPK and ERK MAPK signal transduction. Conclusion: The BFP possessed strong in vitro immune-induced activity, of which fractional purified polysaccharide fractions F1 and F2 were the major immune-induced components. The common signal transduction pathways of BFP, F1, F2 and F3 induced RAW264.7 cells release NO were NF-κB and JNK MAPK. However, the common signal pathways inducing TNF-α secretion in RAW264.7 cells were mainly JNK and ERK MAPK signaling pathways.

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余刚;蔡薇;宋田源;姜泽东;倪辉;刘光明.红毛藻多糖的化学组成及其体外免疫诱导活性研究[J].中国食品学报,2020,20(6):37-47

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