芜菁多糖对巨噬细胞极化及H226细胞增殖和迁移的影响  

Effect of Brassica rapa L.polysachharide on macrophage polarization,proliferation and migration of H226 cells

作　　者：阿曼妮萨·麦提如则 吴美美 李博灵 胡尔沙娜·阿布力克木 迪丽飞拉·苏来曼 海力茜·陶尓大洪 刘九州 MAITIRUZE Amannisa;WU Mei-mei;LI Bo-ling;ABULIKEMU Huershana;SULAIMAN Dilifeila;TAOERDAHON Hailiqian;LIU Jiu-zhou(School of Pharmacy,Xinjiang Medical University;School of Stomatology,Xinjiang Medical University,Urumqi 830017,China)

机构地区：[1]新疆医科大学药学院 [2]新疆医科大学口腔医学院,乌鲁木齐830017

出　　处：《天然产物研究与开发》2025年第1期104-113,共10页Natural Product Research and Development

基　　金：新疆天然药物活性组分与释药技术重点实验室开放课题(2024XJTRY06);新疆维吾尔自治区重大科技专项(2022A03007-3);国家自然科学基金地区科学基金(81960765)。

摘　　要：研究芜菁多糖对巨噬细胞极化以及对H226细胞增殖和迁移的影响。采用CCK-8(cell counting kit-8)法测芜菁粗多糖(Brassica rapa L.crude polysachharide,BRP)、芜菁中性多糖(B.rapa netural polysachharide,BRNP)、芜菁酸性多糖(B.rapa acid polysachharide,BRAP)对RAW 264.7的细胞活力的影响以及BRAP对H226细胞增殖活性的影响;采用线粒体膜电位指示剂检测BRAP对H226细胞线粒体膜电位的变化;用划痕实验检测BRAP对H226细胞的迁移能力的影响。构建M1/M2型巨噬细胞模型,采用酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)检测三种多糖对M1/M2型巨噬细胞的肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)分泌水平的影响,采用免疫荧光法检测三种多糖对M2型巨噬细胞甘露糖受体(mannose receptor,CD206)和一氧化氮合酶(inducible nitric oxide synthase,iNOS)蛋白表达的影响;采用实时荧光定量PCR(realtime fluorescence quantitative PCR,q-PCR)检测三种多糖对M1/M2型巨噬细胞精氨酸酶-1(arginase-1,Arg-1)、白细胞介素-6(interleukin-6,IL-6)的mRNA表达水平的影响。研究结果表明,BRP、BRNP、BRAP对巨噬细胞的增殖起促进作用,并在350μg/mL处对巨噬细胞有较强且相近的促进增殖的作用,BRAP抑制H226细胞增殖效果显著,半数抑制浓度为97.84μg/mL;BRAP降低H226细胞的线粒体膜电位,抑制H226细胞的迁移;三种多糖均能降低巨噬细胞TNF-α及IL-10的分泌量,均能下调M2型巨噬细胞CD206表达,上调iNOS蛋白表达;BRP、BRNP、BRAP降低M1型巨噬细胞IL-6 mRNA的表达以及M2型巨噬细胞Arg-1 mRNA的表达。本研究发现BRP、BRNP、BRAP都能抑制巨噬细胞向M2型表型极化,BRAP能显著抑制H226细胞的增殖和迁移。此研究为芜菁多糖通过调控巨噬细胞极化而抑制肿瘤提供理论基础。To study the effects of Brassica rapa L.polysachharide on macrophage polarization and H226 cell proliferation and migration.The CCK-8(cell counting kit-8)method was used to measure the B.rapa crude polysachharide(BRP),B.rapa netural polysachharide(BRNP),and B.rapa acid polysachharide(BRAP)on the cell viability of RAW 264.7 cells and the effect of BRAP on the proliferative activity of H226 cells.The mitochondrial membrane potential indicator was used to detect the effect of BRAP on the mitochondrial membrane potential of H226 cells.The effect of BRAP on the migration ability of H226 cells was detected by scratch assay.An M1/M2 macrophage model was constructed,and the effects of three polysaccharides on the secretion levels of tumor necrosis factor-α(TNF-α)and interleukin-10(IL-10)in M1/M2 macrophages were detected by enzyme linked immunosorbent assay(ELISA).Immunofluorescence was used to detect the effects of three polysaccharides on the expression of mannose receptor(CD206)and inducible nitric oxide synthase(iNOS)proteins in M2 macrophages.Realtime fluorescence quantitative PCR(q-PCR)was used to detect the effects of three polysaccharides on the expression levels of arginase-1(Arg-1)and interleukin-6(IL-6)in M1/M2 macrophages.The results showed that BRP,BRNP and BRAP promoted the proliferation of macrophages and had a strong and similar effect on macrophages at 350μg/mL,and BRAP had a significant effect on the proliferation of H226 cells,with a half inhibitory concentration of 97.84μg/mL.BRAP decreases the mitochondrial membrane potential of H226 cells and inhibits the migration of H226 cells.The three polysaccharides could reduce the secretion of TNF-α and IL-10 in macrophages,down-regulate the expression of CD206 in M2 macrophages,and up-regulate the expression of iNOS protein.BRP,BRNP and BRAP decreased the expression of IL-6 mRNA in M1 macrophages and Arg-1 mRNA in M2 macrophages.In this study,BRP,BRNP and BRAP could inhibit macrophage polarization to M2 phenotype,and BRAP could significantly inhibit the

关 键 词：芜菁多糖 巨噬细胞 极化 H226细胞 迁移 

分 类 号：R961[医药卫生—药理学]