天然产物研究与开发2024,Vol.36Issue(8) :1288-1297.DOI:10.16333/j.1001-6880.2024.8.002

冬虫夏草多糖的结构表征及改善胞葬功能障碍的作用与机制研究

Structural characterization of Cordyceps polysaccharides and their effects and mechanisms in improving efferocytosis

杨兴茂 杨采霞 周小仝 廖郑悦 刘思静 国锦琳
天然产物研究与开发2024,Vol.36Issue(8) :1288-1297.DOI:10.16333/j.1001-6880.2024.8.002

冬虫夏草多糖的结构表征及改善胞葬功能障碍的作用与机制研究

Structural characterization of Cordyceps polysaccharides and their effects and mechanisms in improving efferocytosis

杨兴茂 1杨采霞 1周小仝 1廖郑悦 2刘思静 2国锦琳3
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作者信息

  • 1. 成都中医药大学药学院现代中药产业学院;西南特色中药资源国家重点实验室
  • 2. 成都中医药大学医学技术学院,成都 610041
  • 3. 成都中医药大学药学院现代中药产业学院;西南特色中药资源国家重点实验室;成都中医药大学医学技术学院,成都 610041
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摘要

基于过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ,PPAR-γ)探究冬虫夏草多糖(Cordyceps polysaccharides,CSP)对改善巨噬细胞胞葬功能的作用及机制.采用水提醇沉法提取CSP;利用高效液相凝胶色谱法、PMP柱前衍生高效液相色谱法、甲基化法分析CSP的分子量分布、单糖组成及糖苷键连接类型.通过CCK-8 法、实时荧光定量 PCR(real-time quantitative polymerase chain reaction,RT-qPCR)和流式细胞术分别测定 CSP对巨噬细胞细胞活力、胞葬相关基因及胞葬功能的影响;采用间接免疫荧光法观察CSP对PPAR-γ的表达量和核定位的影响;利用小干扰RNA转染技术验证PPAR-γ在CSP改善胞葬功能中的作用.结果显示,CSP的总糖含量为72.9%,分子量分布为21.9、2 100 kDa及大于50 000 kDa;主要由葡萄糖、半乳糖和甘露糖三种单糖组成,含6种主要糖苷键.与模型组相比,CSP处理提高了胞葬相关基因TAM酪氨酸激酶受体(Tyro3/Axl/Mer receptor tyrosine ki-nase,TAM)和乳脂球表皮生长因子8(Milk fat globule-epidermal growth factor 8,Mfge8)的表达(P<0.05)以及巨噬细胞对凋亡细胞的胞葬能力(P<0.05).免疫荧光结果表明CSP促进了 PPAR-γ的蛋白表达(P<0.001)与核转位.敲低PPAR-γ后,逆转了 CSP对巨噬细胞胞葬相关基因表达的促进作用.本研究表明CSP可能通过激活PPAR-γ信号通路增强氧化型低密度脂蛋白(Oxidized low-density lipoprotein,ox-LDL)诱导的小鼠巨噬细胞胞葬作用.

Abstract

This study aims to explore the role and mechanism of Cordyceps polysaccharides(CSP)in improving macrophage efferocytosis function based on peroxisome proliferator-activated receptor γ(PPAR-γ).The CSP was extracted by hydro-alco-hol precipitation.The molecular weight distribution,monosaccharide composition and methylation products of CSP were detec-ted by high performance liquid gel chromatography,PMP-high performance liquid chromatography and methylation analysis,respectively.The effects of CSP on mouse macrophage cell viability,efferocytosis-related genes and efferocytosis rate were de-termined by CCK-8 assay,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and flow cytometry,re-spectively.The effect of CSP on PPAR-γ expression and nuclear localization was observed by immunofluorescence.Si-RNA was then used to knock down PPAR-γ to verify the possible target genes and signaling pathways of CSP.The results showed that the CSP contained 72.9%polysaccharides,with a molecular weight distribution of 21.9,2 100 kDa and more than 50000 kDa.The CSP was mainly composed of glucose,galactose and mannose,and it contains six main glycosidic bonds.Com-pared with the model group,CSP increased the expression of efferocytosis-related genes TAM receptor tyrosine kinase(Tyro3/Axl/Mer receptor tyrosine kinase,TAM),milk fat globule-epidermal growth factor 8(Mfge8)and the efferocytosis rate of macrophages(P<0.05).Meanwhile,CSP promoted the protein expression of PPAR-γ(P<0.001)with nuclear transloca-tion.In addition,the ability of CSP to promote the expression of macrophage efferocytosis-related genes was significantly af-fected after knockdown of PPAR-γ(P<0.05).Our study suggests that CSP may enhance oxidized low-density lipoprotein(ox-LDL)-induced macrophage efferocytosis through activating PPAR-γ signaling pathway.

关键词

冬虫夏草/多糖/巨噬细胞/胞葬

Key words

Cordyceps/polysaccharides/macrophages/efferocytosis

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基金项目

国家自然科学基金(81872959)

国家自然科学基金(81373920)

四川省科技厅项目(2023NSFSC1757)

四川省科技厅项目(2023NSFSC3163)

出版年

2024
天然产物研究与开发
中国科学院成都文献情报中心

天然产物研究与开发

CSTPCD北大核心
影响因子:0.783
ISSN:1001-6880
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