连续低强度脉冲式超声联合微泡空化治疗促进小鼠缺血骨骼肌微循环灌注效果的初步评价  

Continuous low-intensity pulsed ultrasound combined with microbubble cavitation therapy promoting microcirculatory perfusion in ischemic skeletal muscle of mice:a preliminary result

作　　者：刘奕佳 汤晓轶 李尚泽 赵佳琦 LIU Yijia;TANG Xiaoyi;LI Shangze;ZHAO Jiaqi(Department of Ultrasound,The Second Affiliated Hospital of Naval Medical University(Second Military Medical University),Shanghai 200003,China;Department of Ultrasound,Shanghai Fourth People’s Hospital Affiliated to Tongji University,Shanghai 200434,China;Department of Orthopedics,Changzhou Medical Branch,No.904 Hospital of Joint Logistics Support Force of PLA,Changzhou 213000,Jiangsu,China)

机构地区：[1]海军军医大学(第二军医大学)第二附属医院超声诊断科,上海200003 [2]同济大学附属上海市第四人民医院超声医学科,上海200434 [3]中国人民解放军联勤保障部队第九〇四医院常州医疗区骨科,常州213000

出　　处：《海军军医大学学报》2025年第3期336-343,共8页Academic Journal of Naval Medical University

基　　金：国家自然科学基金(81501492);上海市自然科学基金(20ZR1457900);上海市虹口区卫生健康委员会医学科研课题面上项目(虹卫2302-26);上海市虹口区卫生健康委员会临床重点扶持专科建设项目(HKLCFC202404);海军军医大学(第二军医大学)第二附属医院人才建设三年行动计划“金字塔人才工程”军事医学人才项目(1009);同济大学附属上海市第四人民医院科研启动专项(SYKYQD06101)。

摘　　要：目的评价连续低强度脉冲式超声(LIPUS)联合微泡(MB)的空化治疗(以下简称超声空化治疗)促进小鼠缺血后肢微循环灌注的效果,初步探讨该治疗方式对肢体动脉缺血性损伤的无创治疗潜力。方法构建小鼠左后肢缺血模型,按照不同治疗方法将小鼠随机分为4组(每组16只):模型组、MB组、LIPUS组和超声空化治疗组(LIPUS+MB组)。模型组小鼠经尾静脉注射0.1 mL生理盐水,MB组小鼠经尾静脉注射0.1 mL MB,LIPUS组小鼠在经尾静脉注射0.1 mL生理盐水的同时采用LIPUS对缺血后肢进行治疗,LIPUS+MB组小鼠在经尾静脉注射0.1 mL MB的同时采用LIPUS对缺血后肢进行治疗;各组均每天注射1次,共7 d。在治疗的第1、4、7天,通过超声造影评价小鼠缺血后肢的微循环灌注情况,并结合小鼠后肢腓肠肌组织H-E染色和CD31免疫组织化学染色,评估不同治疗对小鼠缺血后肢微循环灌注的促进效果。结果小鼠左后肢缺血模型构建成功,所有的模型小鼠均表现出明显的缺血微循环灌注障碍,模型稳定性良好。治疗第7天结束后,LIPUS+MB组小鼠缺血后肢微循环灌注水平升高,其缺血侧与非缺血侧微血管流量比值高于LIPUS组[(94.33±4.51)%vs(70.33±2.09)%,P<0.05];H-E染色结果显示,LIPUS+MB组腓肠肌组织新生毛细血管和肌成纤维细胞数量增加,肌肉结构修复优于LIPUS组,而模型组和MB组则表现为肌细胞坏死、肌束排列紊乱及毛细血管稀疏;CD31免疫组织化学分析进一步证实,超声空化治疗在促进缺血骨骼肌微循环灌注、微血管新生和组织修复方面显著优于单纯LIPUS治疗(CD31相对表达水平5.03±0.33 vs 3.57±0.21,P<0.01)。结论与单纯LIPUS治疗相比,连续超声空化治疗在促进小鼠缺血后肢的微循环灌注方面表现出更加显著的效果,为外周动脉缺血性疾病所致肢体骨骼肌微循环灌注障碍提供了新的治疗策略。Objective To evaluate the promoting effect of continuous low-intensity pulsed ultrasound(LIPUS)combined with microbubble(MB)cavitation therapy(hereinafter referred to as ultrasound cavitation therapy)on microcirculatory perfusion in the ischemic hindlimbs of mice,and to explore the non-invasive therapeutic potential of this treatment for limb arterial ischemic injury.Methods A mouse model of left hindlimb ischemia was established,and the mice were randomly assigned to 4 groups(16 mice per group)according to different treatment methods:model group,ultrasound contrast microbubble group(MB group),LIPUS treatment group(LIPUS group),and ultrasound cavitation therapy group(LIPUS+MB group).Mice in the model group were injected with 0.1 mL of normal saline via the tail vein,those in the MB group were injected with 0.1 mL of MB via the tail vein,those in the LIPUS group were treated with LIPUS on the ischemic hindlimb after injection of 0.1 mL of normal saline via the tail vein,and those in the LIPUS+MB group were treated with LIPUS on the ischemic hindlimb after injection of 0.1 mL of MB via the tail vein;each group was injected once a day for a total of 7 d.On the 1st,4th and 7th days after treatment,the microcirculatory perfusion in the ischemic hindlimbs of mice was evaluated using contrast-enhanced ultrasound.The effects of different treatments on promoting microcirculatory perfusion in the ischemic hindlimbs of mice were assessed by combining hematoxylin-eosin(H-E)staining and CD31 immunohistochemical staining of the gastrocnemius muscle tissue in the hindlimbs.Results The left hindlimb ischemia model was successfully constructed,and all model mice showed obvious ischemic microcirculation perfusion disorders with good model stability.After the 7th day of treatment,the LIPUS+MB group showed a increase in microcirculation perfusion in the ischemic hindlimb,with the ratio of microvascular flow on the ischemic to non-ischemic sides higher than that of the LIPUS group([94.33±4.51]%vs[70.33±2.09]%,P<0.05).H-E staining r

关 键 词：微泡空化治疗 低强度脉冲式超声波治疗 超声造影 微循环灌注 外周动脉疾病 

分 类 号：R543.5[医药卫生—心血管疾病]