摘要
目的:研究马尾藻多糖(SP)对抗脂多糖(LPS)诱导的巨噬细胞极化和铁死亡,以明确SP是否存在对免疫功能的调节作用。方法:分离、提纯和鉴定SP,建立LPS诱导的小鼠巨噬细胞RAW264.7炎症模型。采用CCK-8法检测SP对巨噬细胞增殖的影响,采用实时荧光定量PCR (RT-qPCR)法和Western blot法分别检测巨噬细胞极化相关指标CD80、CD163、IL-6、IL-10、精氨酸-1(Arg-1)、诱导型一氧化氮合酶(iNOS)、TNF-α含量。为了解巨噬细胞铁死亡情况,以活性氧(ROS)检测试剂盒检测ROS含量,并以Western blot法检测谷胱甘肽过氧化物4(GPX4)的表达。结果:成功分离SP,并鉴定其不属于呋喃糖。高浓度SP对巨噬细胞具有促增殖作用,并对LPS诱导的细胞增殖抑制具有显著的保护作用。SP能使RAW264.7细胞向M2极化,并且能拮抗LPS诱导的M1极化。LPS可诱导巨噬细胞内ROS显著增加,SP则可使其ROS水平显著降低。LPS诱导巨噬细胞的GPX4蛋白表达水平显著降低,高浓度的SP能显著上调其表达(P均<0.05)。结论:SP诱导巨噬细胞向M2极化,拮抗LPS诱导的巨噬细胞M1型极化和铁死亡。
Objective:To study the protective effect of sargassum polysaccharides(SP)on macrophage polarization and iron death induced by lipopolysaccharide(LPS)in order to determine whether SP has a regulatory effect on immune function.Methods:Isolating,purificating and identificating SP,a mouse macrophage RAW264.7 cell line inflammation model induced by LPS was established.The CCK-8 method was used to detect the effect of SP on macrophage proliferation.The levels of CD80,CD163,IL-6,IL-10,Arginine-1(Arg-1),inducible nitric oxide synthase(iNOS),IL-6 and TNF-αin macrophages were measured by real-time fluorescence quantitative PCR(RT-qPCR)and Western blot.In order to understand the iron death of macrophages,ROS content was detected with ROS assay kit,and glutathione peroxid 4(GPX4)expression was detected by Western blot.Results:SP were isolated successfully and identified not as furanose.High concentration of SP promote the proliferation of macrophages and had a significant protective effect on the inhibition of cell proliferation induced by LPS.SP could polarize RAW264.7 cells to M2 and antagonize M1 polarization induced by LPS.LPS could significantly increase ROS in macrophages,while SP could significantly decrease the level of ROS.The expression of GPX4 protein in macrophages induced by LPS was significantly decreased,and the expression was significantly up-regulated by high concentration of SP(P<0.05).Conclusion:SP induces macrophages M2 polarization and antagonizes LPS induced M1 polarization and ferroptosis of macrophages.
作者
袁慧情
胡佳敏
刘思溢
侯鉴基
吴科锋
罗辉
吴斌华(指导)
YUAN Huiqing;HU Jiamin;LIU Siyi;HOU Jianji;WU Kefeng;LUO Hui;WU Binhua(Guangdong Medical University,Zhanjiang 524023,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第10期2034-2040,共7页
Chinese Journal of Immunology
基金
广东省医学科研基金项目(A2023207)
广东省中医药局中医药科研项目(20242043)。
关键词
马尾藻多糖
巨噬细胞
细胞极化
铁死亡
Sargassum polysaccharides
Macrophages
Cell polarization
Ferroptosis
