人参外泌体通过促进巨噬细胞M1极化抑制肺癌细胞生长  

Ginseng-derived Nanoparticles Inhibit Lung Cancer Cell Growth by Promoting Macrophage M1 Polarization

作　　者：范亮亮 任李梅 杨松 李文婧 赵悦名 赵荣华 赵大庆[2] 王佳雯[2] FAN Liangliang;REN Limei;YANG Song;LI Wenjing;ZHAO Yueming;ZHAO Ronghua;ZHAO Daqing;WANG Jiawen(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China;Northeast Asia Institute of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学吉林省人参科学研究院,吉林长春130117 [2]长春中医药大学东北亚中医药研究院,吉林长春130117

出　　处：《食品工业科技》2025年第4期358-366,共9页Science and Technology of Food Industry

基　　金：吉林省教育厅科学技术研究项目(NO.JJKH20220875KJ)。

摘　　要：目的:旨在探讨人参外泌体(Ginseng-derived nanoparticles,GDNPs)通过改变巨噬细胞极化状态,抑制人非小细胞肺癌细胞(A549)生长的分子机制,为人参鲜药的深入研究提供一定的理论基础。方法:采用CCK-8法检测人参外泌体对巨噬细胞(RAW264.7)活力的影响;实时定量PCR检测人参外泌体刺激巨噬细胞M1极化相关指标IL-6、iNOS、TNF-α、MCP-1转录水平;流式细胞术检测M1型巨噬细胞分子标志CD80的表达。收集LPS和人参外泌体处理的RAW264.7细胞衍生的上清液,制成条件培养基(CM),并与A549共培养。CCK8和流式细胞术检测A549细胞活力、细胞周期和细胞凋亡;采用Western blot检测GDNPs-CM对A549细胞中TLR4/MyD88通路相关蛋白表达的影响。结果:人参外泌体能够增强巨噬细胞的细胞活力(P<0.05,P<0.01),并提高巨噬细胞M1极化相关指标IL-6、iNOS、TNF-α、MCP-1转录水平的表达(P<0.05,P<0.01,P<0.0001);人参外泌体增强了M1型巨噬细胞分子标志CD80的表达(P<0.01)。在条件培养基作用下,A549细胞的细胞活力受到抑制,并发生G1期阻滞,A549细胞的凋亡率提高(P<0.05,P<0.01);同时,GDNPs-CM促进了A549细胞中TLR4、MyD88、NF-κB、iNOS、COX-2等炎症相关蛋白表达量(P<0.05,P<0.01)。结论:人参外泌体通过刺激巨噬细胞M1极化,抑制A549细胞增殖,调节A549细胞周期,激活TLR4/MyD88/NF-κB信号通路提高炎症因子的表达水平,从而诱导肺癌细胞凋亡。Objective:To explore the molecular mechanism by which ginseng-derived nanoparticles(GDNPs)inhibit the growth of human non-small cell lung cancer cells(A549)by altering the polarization state of macrophages.This study will provide a theoretical basis for further research on fresh ginseng medicine.Methods:The CCK-8 assay was employed to detect the effect of GDNPs on the viability of macrophages(RAW264.7).Real-time quantitative PCR was utilized to detect the transcript levels of IL-6,iNOS,TNF-α,and MCP-1,which were indicators related to the M1 polarization of macrophages stimulated by GDNPs.Flow cytometry was utilized to detect the expression of M1 macrophages molecular markers CD80,which was related to the M1 polarization of macrophages stimulated by GDNPs.RAW264.7 cells were treated with LPS and GDNPs,and the culture supernatants were collected to prepare a conditional medium(CM),which was subsequently co-cultured with A549 cells.CCK-8 and flow cytometry were employed to assess the cell viability,cell cycle,and apoptosis of A549 cells.Western blotting was performed to investigate the effect of GDNPs-CM on the expression of TLR4/MyD88 pathway-related proteins in A549 cells.Results:GDNPs enhanced the cell viability of macrophages(P<0.05,P<0.01)and increased the transcript levels of IL-6,iNOS,TNF-α,and MCP-1(P<0.05,P<0.01,P<0.0001).GDNPs enhanced the the expression of M1 macrophages molecular markers CD80(P<0.01).Under the influence of GDNPs-CM,the cell viability of A549 cells was inhibited,and the cells exhibited G1 phase arrest with an increased apoptosis rate(P<0.05,P<0.01).Simultaneously,GDNPs-CM promoted the expression levels of inflammationrelated proteins,such as TLR4,MyD88,NF-κB,iNOS,and COX-2 in A549 cells(P<0.05,P<0.01).Conclusion:GDNPs induce apoptosis in lung cancer cells by stimulating M1 polarization of macrophages,inhibiting the proliferation of A549 cells,regulating the A549 cell cycle,and activating the TLR4/MyD88/NF-κB signaling pathway to promote the expression levels of inflammatory factors.

关 键 词：人参外泌体 肿瘤相关巨噬细胞 人非小细胞肺癌细胞 细胞周期 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]