膜联蛋白V纳米微泡抑制ox-LDL诱导的巨噬细胞组织因子表达的实验研究  

Experimental Study on Inhibition of Tissue Factor Expression in ox-LDL-induced Macrophages by Annexin V Nanobubbles

作　　者：赵萍 马晓菊 陈建梅 袁丽君 段云友 Zhao Ping;Ma Xiaoju;Chen Jianmei;Yuan Lijun;Duan Yunyou(Department of Ultrasonic Medicine,Tangdu Hospital,Air force Medical University,Xi’an,Shannxi 710038.China)

机构地区：[1]空军军医大学唐都医院超声医学科,西安市710038

出　　处：《中国超声医学杂志》2021年第10期1182-1185,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金青年项目(No.81201136);空军军医大学唐都医院苗子人才资助基金(No.2019MZRC001)。

摘　　要：目的分析膜联蛋白V (AV)纳米微泡对组织因子表达的影响及机制,探讨其作为抗凝治疗分子探针的可能性。方法巨噬细胞RAW 264.7分别给予氧化型低密度脂蛋白(ox-LDL)和生理盐水诱导12 h以上,流式细胞仪检测各组细胞凋亡率;以"优化的薄膜水化"和"生物素-亲和素"法制备AV纳泡,激光共聚焦观察其与ox-LDL诱导的巨噬细胞靶向结合能力;单纯ox-LDL和ox-LDL+AV纳泡分别诱导细胞0、6、12、18 h和24 h,酶标仪检测细胞上清液中TF含量,统计学分析差异显著性。结果 ox-LDL诱导12 h以上,细胞凋亡率达(59.74±1.67)%,与对照组(1.23±0.29)%差异显著(P<0.05);大量AV纳泡聚集在ox-LDL诱导的巨噬细胞周围;单纯ox-LDL可诱导细胞高表达TF,AV纳泡与ox-LDL同时加入后,TF显著降低,统计学分析加AV纳泡前后数据具有显著性差异(P<0.05)。结论 AV纳泡靶向结合ox-LDL诱导的巨噬细胞可能抑制了TF的表达,提示其潜在的抗凝价值。Objective To investigate the effect and mechanism of annexin V-nanobubbles(AV-nbs) on tissue factor(TF), and explore the possibility of AV-nbs as a potential molecular probe for anticoagulant therapy. Methods The RAW 264.7 cell apoptosis rate were measured by flow cytometry after oxidized low-density lipoprotein(ox-LDL) and saline treatment for 12 h. The AV-nbs were prepared by “optimized film hydration” and “biotin-avidin-biotin” method. The ability of AV-nbs binding to macrophages was determined via the laser scanning confocal microscope. Macrophages were induced by ox-LDL and ox-LDL+AV for 0, 6, 12, 28 and 24 h, respectively. Then, the TF was detected by enzyme linked immunosorbent assay(ELISA) and the significant difference was analyzed by statistical analysis. Results The apoptotic proportion of ox-LDL group was(59.74±1.67)%, compared with(1.23±0.29)% in control(P<0.05). A large number of AV-nbs accumulated on the surface of ox-LDL-induced macrophages. The level of TF increased by ox-LDL, but declined by AV-nbs(P<0.05). Conclusions The TF was inhibited by AV-nbs probably via targeting apoptotic macrophage, suggesting its potential value for anticoagulant therapy.

关 键 词：巨噬细胞 组织因子 细胞凋亡 Annexin V 纳米微泡 

分 类 号：R543.5[医药卫生—心血管疾病]