菊粉对巨噬细胞RAW 264.7免疫活性的影响  

Effect of inulin on the immune activity of RAW 264.7 macrophages

作　　者：牛甜 杨帅 段宏伟 成文婧 张丽鸿 胡俊杰[1] NIU Tian;YANG Shuai;DUAN Hong-wei;CHENG Wen-jin;ZHANG Li-hong;HU Jun-jie(Gansu Provincial Key Laboratory of Animal Reproductive Physiology and Reproductive Regulation,College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院甘肃省动物生殖生理及繁殖调控重点实验室,兰州730070

出　　处：《天然产物研究与开发》2025年第1期10-17,共8页Natural Product Research and Development

基　　金：甘肃农业大学伏羲青年英才项目(Gaufx-05Y07);甘肃农业大学公招博士科研启动基金(GAU-KYQD-2021-09)。

摘　　要：菊粉(inulin,INU)作为一种植物多糖,因具有广泛的应用价值以及增强免疫、抗炎、抗肿瘤等生物学功能而备受关注,本研究旨在探究INU对巨噬细胞RAW 264.7免疫活性的影响。采用细胞增殖检测试剂盒(cell counting kit-8,CCK8)、中性红试验检测INU对巨噬细胞RAW 264.7增殖活性和吞噬活性的影响。采用格里斯试剂法和免疫酶联吸附试验(enzyme-linked immunosorbent assay,ELISA)检测细胞中一氧化氮(nitric oxide,NO)和细胞因子白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的释放量。实时荧光定量PCR(real-time fluorescence quantitative PCR,qPCR)检测细胞因子(IL-1β、IL-6、TNF-α)的基因表达量以及免疫蛋白印迹(Western blotting,WB)检测INU对巨噬细胞RAW 264.7中环氧化酶2(cyclooxygenase 2,COX2)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和丝裂原蛋白活化激酶(mitogen activated protein kinase,MAPK)相关通路关键蛋白表达的影响。实验结果表明,INU显著增强RAW 264.7细胞的增殖活性、吞噬活性并促进NO的分泌及IL-1β、IL-6、TNF-α的表达水平(P<0.01)。此外,INU还以浓度依赖的方式显著上调了COX2、iNOS及MAPK通路中磷酸化细胞外调节蛋白激酶(phosphorylated extracellular regulatory protein kinase,p-ERK)、磷酸化p38丝裂原活化蛋白激酶(phosphorylated p38 mitogen-activated protein kinase,p-p38)的蛋白表达(P<0.01)。综上所述,INU可通过ERK/p38MAPK通路调控巨噬细胞的免疫活性,具有良好的免疫调节能力。Inulin(INU),as a plant polysaccharide,has attracted much attention due to its wide application value and enhancement of immune,anti-inflammatory,anti-tumor and other biological functions.The aim of this study was to investigate the effect of INU on the immune activity of macrophage RAW 264.7.The effects of INU on the proliferative activity and phagocytotic activity of RAW 264.7 macrophages were detected by cell counting kit-8(CCK8)and neutral red test.The Griess assay and enzyme-linked immunosorbent assay(ELISA)were used to detect the release of nitric oxide(NO)and cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in cells.Real time fluorescence quantitative PCR(qPCR)was used to detect the gene expression levels of cytokines(IL-1β,IL-6,TNF-α),and Western blotting(WB)was used to detect the effects of INU on the expression of key proteins related to cyclooxygenase 2(COX2),inducible nitric oxide synthase(iNOS),and mitogen activated protein kinase(MAPK)pathways in RAW 264.7 macrophages.The results showed that INU significantly enhanced the proliferative activity and phagocytosis activity of RAW 264.7 cells and promoted the secretion of NO and the expression levels of IL-1β,IL-6 and TNF-α(P<0.01).In addition,INU significantly up-regulated the protein expression of phosphorylated extracellular regulatory protein kinase(p-ERK)and phosphorylated p38 mitogen-activated protein kinase(p-p38)in COX2,iNOS and MAPK pathways in a concentration-dependent manner(P<0.01).In summary,INU can regulate the immune activity of macrophages through the ERK/p38MAPK pathway,and has a good ability of immune regulation.

关 键 词：菊粉 巨噬细胞RAW 264.7 MAPK信号通路 免疫活性 

分 类 号：R967[医药卫生—药理学]