氧化铁纳米颗粒在Caco-2细胞模型,上的吸收机制  

Absorption mechanism of iron oxide nanoparticles in Caco-2 cell model

作　　者：苏欣怡 赵靖 刘洋 侍江春 谢羽檬 罗雅亭 李云 Su Xinyi;Zhao Jing;Liu Yang;Shi Jiangchun;Xie Yumeng;Luo Yating;Li Yun(West China School of Public Health,Sichuan University,Chengdu 610041,China;Academy of Medical Sciences Department of Nurtriology,Sichuan Provincial People's Hospital,Chengdu 610072,China)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院,成都610041 [2]四川省医学科学院.四川省人民医院营养科,成都610072

出　　处：《卫生研究》2024年第6期992-998,共7页Journal of Hygiene Research

基　　金：国家重点研发计划(No.2017YFC1600204)。

摘　　要：目的探究氧化铁纳米颗粒(Fe_(2)O_(3)NPs)经胃肠道吸收进入人体的可能机制。方法以Caco-2单层细胞为细胞模型,将已表征的Fe_(2)O_(3)NPs制备为悬液后干预Caco-2细胞,采用CCK-8法、Transwell法、原子分光光度计法探究Fe_(2)O_(3)NPs对Caco-2细胞活性的影响以及经Caco-2单层细胞模型的吸收转运情况。采用不同剂量的Fe_(2)O_(3)NPs悬液处理Caco-2细胞后,通过电镜观察、内吞抑制剂(氯丙嗪、甲基-β-环糊精、阿米洛利)及蛋白免疫印迹法探究Fe_(2)O_(3)NPs在Caco-2细胞上的吸收机制。结果根据细胞活性、吸收转运试验结果选择1.5、1.0、0.5 mg/mL作为吸收机制研究暴露剂量。电镜下显示细胞内外均分布Fe_(2)O_(3)NPs,细胞内的Fe_(2)O_(3)NPs存在于细胞质内和囊泡样结构中,内吞抑制剂氯丙嗪和甲基-β-环糊精处理24 h对Fe_(2)O_(3),NPs在Caco-2细胞模型上的吸收均表现出抑制作用。Fe_(2)O_(3)NPs处理Caco-2细胞能使细胞二价金属转运体蛋白、膜铁转运蛋白表达量上调(P<0.01),闭锁蛋白表达量下调(P<0.05)。结论Fe_(2)O_(3)NPs在Caco-2细胞上存在3种吸收机制:网格蛋白和小窝/脂筏蛋白介导的内吞途径、铁转运相关蛋白参与的吸收转运、伴随紧密连接蛋白破坏的细胞旁途径。OBJECTIVE To explore the possible mechanism of absorption of iron oxide nanoparticles into the human body through the gastrointestinal tract.METHODS This article used Caco-2 monolayer cells as a cell model,prepared characterized iron oxide nanoparticles(Fe_(2)O_(3)NPs)as suspensions,and intervened in Caco-2 cells.CCK-8 method,transwell method,and atomic spectrophotometer method were used to explore the effect of Fe_(2)O_(3)NPs on the activity of Caco-2 cells and the absorption and transport of them through the Caco-2 monolayer cell model.After treating Caco-2 cells with different doses of iron oxide nanoparticles suspension,electron microscopy observation and endocytosis inhibitorss(chlorpromazine,methyl-β-cyclodextrin,amiloride)and western blot were performed exploring the absorption mechanism of iron oxide nanoparticles on Caco-2 cells.RESULTS1.5,1.0 and 0.5 mg/mL were selected as absorption mechanism studies and exposure doses based on cell activity and absorption transport test result.Under electron microscopy,Fe_(2)O_(3)NPs were distributed both inside and outside the cell.Fe_(2)O_(3)NPs presented in the cytoplasm and vesicular like structures within cells.Endocytosis inhibitors chlorpromazine and methyl-β-cyclodextrin treatment for 24 hours showed inhibitory effects on the absorption ofFe_(2)O_(3)NPs in the Caco-2 cell model.Treatment of Caco-2 cells with Fe_(2)O_(3)NPs could increase the expression of divalent metal transporter(DMT1)and ferroportin 1(FPN1)(P<0.01),while decreasing expression of occludin(P<0.05).CONCLUSION There are three absorption mechanisms of iron oxide nanoparticles on Caco-2 cells:endocytosis pathways mediated by reticulin and caveolin/lipid raft proteins,absorption and transport involving iron transport related proteins,and paracellular pathways accompanied by the disruption of tight junction proteins.

关 键 词：氧化铁纳米颗粒 CACO-2细胞 吸收机制 

分 类 号：R155.5[医药卫生—营养与食品卫生学]