超声纳泡靶向抑制IRF5调控易损斑块巨噬细胞极化的实验研究  

Ultrasound-Sensitive IRF5-Targeting siRNA-loaded Nanobubble on the Regulation of Macrophage Polarization in Vulnerable Plaques

作　　者：姚玉娟 王宇豪 李妙[1] 方玲玲[1] 万林林[1] 李林[1] 张平洋[1] Yao Yujuan;Wang Yuhao;Li Miao;Fang Lingling;Wan Linlin;Li Lin;Zhang Pingyang(Department of Cardiovascular Ultrasound,Nanjing First Hospital,Nanjing Medical University,Nanjing 21o006,China)

机构地区：[1]南京医科大学附属南京医院(南京市第一医院)心血管超声科,南京市210006

出　　处：《中国超声医学杂志》2023年第11期1293-1296,共4页Chinese Journal of Ultrasound in Medicine

基　　金：江苏省卫生健康委重点医学科研项目(No.ZD2021048)。

摘　　要：目的探讨携IRF5-siRNA的靶向超声纳泡在超声作用下介导基因转染,对易损斑块M1型巨噬细胞极化的靶向调控作用。方法构建携IRF5-siRNA靶向超声纳泡,检测其基本表征并观察其靶向结合能力。建立M1型巨噬细胞,将其分为PBS组、IRF5-siRNA质粒组、IRF5-siRNA-PP1-NBs组、US+IRF5-siRNA-PP1-NBs组。qRT-PCR法检测各组IRF5 mRNA表达;Western bolt(blot)法检测各组iNOS、Arg1蛋白表达;流式细胞术检测巨噬细胞亚型。结果制得的超声纳泡平均粒径为(304.48±59.34)nm,Zeta电位为(32.75±4.02)mV;大量纳泡聚集在M1型巨噬细胞周围;与其他组相比,US+IRF5-siRNA-PP1-NBs组IRF5 mRNA表达显著降低,M1型巨噬细胞标志物iNOS、CD86表达降低,M2型巨噬细胞标志物Arg1、CD206表达增高。结论成功构建了携IRF5-siRNA靶向超声纳泡,在超声介导下可有效实现M1型巨噬细胞基因转染并调控其表型,提示其潜在的促进斑块稳定性价值。Objective To investigate the targeted regulation of the polarization of Ml macrophages in vulnerable plaque by ultrasound-sensitive IRF5-targeting siRNA-loaded nanobubbles.Methods We Constructed IRF5-targeting siRNA-loaded nanobubbles,examined their basic characterization,and observed their targeted binding ability.We then established M1 macrophages,and divided them into four groups:PBS group,IRF5-siRNA plasmid group,IRF5-siRNA-PP1-NBs group,and US+IRF5-siRNA-PP1-NBs group.qRT-PCR was used to detect the expression of IRF5 mRNA in each group.Western blotting was performed to evaluate the protein expression of iNOS and Argl in each group.Flow cytometry was employed to examine macrophage subtypes.Results The prepared IRF5-siRNA-PP1-NBs exhibited an average particle size of(304.48±59.34)nm and a Zeta potential of(32.75±4.02)mV.A substantial accumulation of nanobubbles was observed surrounding Ml-type macrophages.In comparison to the other experimental groups,the US+IRF5-siRNA-PP1-NBs group exhibited a significant reduction in IRF5 mRNA expression,accompanied by decreased expression levels of Ml macrophage markers,including iNOS and CD86.Conversely,the expression of M2 macrophage markers,including Argl and CD206,was found to be elevated.Conclusions Ultrasoundsensitive IRF5-targeting siRNA-loaded nanobubbles are successfully constructed which effectively achieved gene transfection in Ml macrophages and regulated their phenotype under ultrasound intervention.These findings suggest the potential value of these nanobubbles in promoting plaque stability.

关 键 词：巨噬细胞极化 易损斑块 靶向超声 纳泡 

分 类 号：R54[医药卫生—心血管疾病] R454[医药卫生—内科学]