果糖通过促进糖酵解调控M1型巨噬细胞极化  

Fructose regulates M1 macrophage polarisation by promoting glycolysis

作　　者：王名鸿 刘彧冰 王新航 李菡 陈思宏 张敏华 彭阳 陆彩玲 唐深[2,3] 李习艺[1] WANG Minghong;LIU Yubing;WANG Xinhang;LI Han;CHEN Sihong;ZHANG Minhua;PENG Yang;LU Cailing;TANG Shen;LI Xiyi(School of Public Health,Guangxi Medical University,Nanning 530021,China;School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021,China;Key Laboratory of Basic Research on Regional Diseases,Education Department of Guangxi Zhuang Autonomous Region,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学公共卫生学院,南宁530021 [2]广西医科大学基础医学院,南宁530021 [3]广西医科大学广西高校区域性疾病基础研究重点实验室,南宁530021

出　　处：《广西医科大学学报》2025年第2期168-173,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82460638,No.82160612)。

摘　　要：目的:探讨果糖(Fru)对脂多糖(LPS)和干扰素γ(IFN-γ)诱导的M1型巨噬细胞极化过程的调控作用及可能机制。方法:实验分为M0组(THP-1细胞用100 nmol/L佛波酯刺激24 h)、M1组(M0组基础上用10 ng/mL LPS和20 ng/mL IFN-γ刺激48 h)、M1+5 mmol/L Fru组(加入LPS和IFN-γ的同时加入5 mmol/L Fru刺激48 h)。实时荧光定量PCR(RT-qPCR)检测M1型巨噬细胞极化标志物白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和环氧化物酶-2(COX-2)的mRNA表达;微量法酶活性试剂盒检测糖酵解限速酶己糖激酶(HK)、乳酸脱氢酶(LDH)活性;活性氧(ROS)试剂盒检测细胞内ROS水平;中性红吞噬实验检测细胞吞噬能力;刃天青法检测线粒体呼吸链代谢酶活性;三磷酸腺苷(ATP)检测试剂盒检测ATP含量。结果:与M0组比较,M1组IL-1β、IL-6、TNF-α和COX-2 mRNA表达增加,HK、LDH活性升高,线粒体呼吸链代谢酶活性和ATP含量降低,细胞内ROS生成增加,吞噬能力增强(均P<0.05)。与M1组比较,M1+5 mmol/L Fru组IL-1β、IL-6、TNF-α和COX-2 mRNA表达水平升高,HK、LDH活性增加,线粒体呼吸链代谢酶活性和ATP含量降低,细胞内ROS生成增加,吞噬能力降低(均P<0.05)。结论:Fru可能通过促进M1型巨噬细胞糖酵解,降低氧化磷酸化水平,上调M1型巨噬细胞极化相关标志物的表达,促进ROS产生,下调巨噬细胞吞噬能力,从而调控M1型巨噬细胞极化。Objective:To investigate the regulatory effect of fructose(Fru)on the polarization of M1 macrophages induced by lipopolysaccharide(LPS)and interferon gamma(IFN-γ)and its possible mechanism.Methods:The experiment was divided into M0 group(THP-1 cells were stimulated with 100 nmol/L phorbol esters for 24 h),M1 group(M0 group was stimulated with 10 ng/mL LPS and 20 ng/mL IFN-γfor 48 h),and M1+5 mmol/L Fru group(5 mmol/L Fru was added simultaneously with LPS and IFN-γfor 48 h of stimulation).Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression of polarization markers interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and cyclooxygenase-2(COX-2)in M1 macrophages.The activities of glycolytic rate-limiting enzyme hexokinase(HK)and lactate dehydrogenase(LDH)were detected using a microenzyme activity kit.The reactive oxygen species(ROS)kits were used to detect intracellular ROS levels.The phagocytic ability of the cells was detected by neutral erythrophagocytosis assay.The mitochondrial respiratory chain metabolic enzyme activities were detected by Azulin method.The adenosine triPhosphate(ATP)contents were detected by using an ATP test kit.Results:Compared with the M0 group,mRNA expression of IL-1β,IL-6,TNF-αand COX-2 in the M1 group was increased,activities of HK and LDH were increased,mitochondrial respiratory chain metabolic enzyme activities and ATP contents were decreased,intracellular ROS production was increased,and phagocytic ability was enhanced(all P<0.05).Compared with the M1 group,mRNA expression levels of IL-1β,IL-6,TNF-αand COX-2 in the M1+5 mmol/L Fru group were increased,activities of HK and LDH were increased,mitochondrial respiratory chain metabolic enzyme activities and ATP contents were decreased,intracellular ROS production was increased,and phagocytic ability was decreased(all P<0.05).Conclusion:Fructose may promote the glycolysis of M1 macrophages,reduce the level of oxidative phosphorylation,upregulate the expres

关 键 词：果糖 M1型巨噬细胞 糖酵解 极化 吞噬能力 

分 类 号：R151[医药卫生—营养与食品卫生学]