靶向抗ICAM-1的载伊马替尼和液态氟碳脂质体对内毒素诱导的急性肺损伤/急性呼吸窘迫综合征模型小鼠的治疗研究  

Anti-ICAM targeted liposomes loaded with imatinib and liquid fluorocarbon treat ALI/ARDS mouse model induced by LPS

作　　者：胡聪[1] 李攀[2] 王志刚[2] 江德鹏[1] HU Gong;LI Pan;WANG Zhigang;JIANG Depeng(Department of Respiratory Diseases,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010;Chongqing Key Laboratory of Ultrasound Molecular Imaging,Chongqing Medical University,Chongqing,400010,China)

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010 [2]重庆医科大学超声分子影像重庆市重点实验室,重庆400010

出　　处：《第三军医大学学报》2018年第15期1357-1363,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金应急管理项目(81650003)~~

摘　　要：目的探讨靶向抗细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)的载伊马替尼(imatinib mesylate,IM)和液态氟碳(liquid fluorocarbon,LF)脂质体对内毒素(lipopolysaccharides,LPS)诱导的急性肺损伤/急性呼吸窘迫综合征(acute lung injury/acute respiratory distress syndrome,ALI/ARDS)模型小鼠的治疗效果。方法用旋转蒸发法制备靶向抗ICAM-1的载伊马替尼和液态氟碳脂质体,用马尔文粒径仪器检测其粒径,透射电镜下观察脂质体内部载药情况。光镜下观察其形态,置于37.5℃恒温加热板上观察其相变情况。体外模拟体内药物释放环境,观察脂质体内包载的伊马替尼药物的释放。6~8周大小30只C57BL/6小鼠按随机数字表法分为:对照组,LPS造模组,靶向抗ICAM-1脂质体治疗组,每组10只。治疗结束1 d后观察肺石蜡切片HE染色和肺石蜡切片免疫荧光ICAM-1的分布情况,肺组织提取mRNA,用qRT-PCR检测IL-6,IL-8,IL-10,TNF-α表达情况,同时对比肺的病理切片,小鼠肺湿干重比和小鼠肺泡液体清除率。靶向抗ICAM-1脂质体和非靶向脂质体分别尾静脉注射入ALI/ARDS小鼠体内,各个器官做冰冻切片,共聚焦激光显微镜下观察两组脂质体在小鼠体内各个器官的分布。结果成功制备靶向抗ICAM-1的载伊马替尼和液态氟碳脂质体,其粒径为(320.8±58.7)nm,透射电镜下观察其内部成功包载药物,光镜下看到脂质体大小均匀,在37.5℃恒温板发生相变形成微泡。12 h脂质体药物释放达到高峰,绘出脂质体药物的释放曲线。与对照组相比,LPS造模组有典型的ALI/ARDS病理特征,肺损伤明显,病理切片评分增加(P<0.01),炎症因子IL-6,IL-8,TNF-α增加(P<0.01),IL-10减少(P<0.05)。湿干重比(W/D)增高(P<0.01),而小鼠肺泡液体清除率(alveolar fluid clearance,AFC)明显减弱(P<0.01)。靶向抗ICAM-1脂质体治疗组肺损伤减缓,病理切片评分下降(P<0.05),炎症因子IL-6,IL-8,TNF-α减少(P<0.05),IL-10增加(PObjective To determine the therapeutic effect of intercellular cell adhesion molecule-1 （ICAM-1） targeted liposomes loaded with imatinib （IM） and liquid fluorocarbon （LF） on the treatment of lipopolysaccharides （LPS）-induced mouse model of acute lung injury/acute respiratory distress syndrome （ALI/ARDS）. Methods The anti-ICAM-1 liposomes were prepared by rotary evaporation to load IM and LF, and the particle size was detected by Malvin particle size analyzer. Transmission electron microscopy was used to observe the loading of drugs in the liposomes, while under light microscopy for the shape of the prepared liposomes. Phase transformation was observed at constant temperature heating plate at 37.5 ℃, and release rate of the drugs from the liposomes was observed in vitro. A total of 30, 6 - 8-week-old C57BL/6 mice were randomly divided into control group, LPS model group, and targeting ICAM-1 liposomes treatment group （n = 10）. In 1 d after the end of treatment, HE staining and immunofluorescence assay were employed to observe the morphology and distribution of ICAM-1 in the lung paraffin sections respectively. The mRNA levels of IL-6, IL-8, IL-10 and TNF-α in the lung tissue were detected by qRT-PCR. Pathological changes, wet-to-dry （W/D） lung weight ratio, and alveolar fluid clearance （AFC） were compared in the different groups. ICAM-1 targeted liposomes and non-targeted liposomes were injected into ALI/ARDS mice by tail vein. All organs were collected for frozen sections, and the distributions of the liposomes in various organs were observed under confocal laser microscope. Results ICAM-1 targeted liposomes loaded with IM and LF were successfully prepared, with a particle size of 320. 8 ± 58. 7 nm. Transmission electron microscopy displayed the drugs were successfully encapsulated inside them, while light microscopy showed the liposomes were uniform in size, and gradually formed into microbubbles at 37.5 ℃. The release of liposomes reached peak at 12 h, and the release curve

关 键 词：急性肺损伤 急性呼吸窘迫综合征 伊马替尼 脂质体 细胞间黏附分子-1 

分 类 号：R-332[医药卫生] R563.8