诊断超声介导自制微泡造影剂联合尿激酶体外溶栓效应及参数优化的实验研究  

Effect of Diagnostic Ultrasound-mediated Microbubble Contrast and Urokinase on Augmentation Thrombolysis and Optimization of the Major Parameters:an in vitro Study

作　　者：喻琴梅[1] 王红[2] 孟文彤[1] 李艳秋[1] 杨金荣[1] 李军[1] 周翔[2] 牛挺[1] 

机构地区：[1]四川大学华西医院血液科血液学研究室,成都610041 [2]四川大学华西医院超声科,成都610041

出　　处：《四川大学学报（医学版）》2012年第3期451-457,共7页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30970839);四川省科技厅-应用基础(批准号2010JY0006)资助

摘　　要：目的研究诊断超声介导自制微泡造影剂对体外血栓的溶解效应及探讨影响溶栓效果的主要因素,确定最优溶栓参数组合。方法制备标准化红色血栓,设计、建立体外溶栓模型。选用L16(45)正交表设计不同输出能量(因素A:5%、25%、50%、100%)、不同微泡量(因素B:50、100、200、400μL)、不同尿激酶浓度(因素C:100、200、400、800U/mL)、不同溶栓时间(因素D:10、20、30、40min)的4因素4水平实验组合,在固定诊断超声频率为1.82MHz条件下,按照组合对制备好的4种栓龄(3、6、12、24h)标准红色血栓进行体外溶栓实验。采用组织病理切片HE染色及扫描电镜观察溶栓前后血栓结构。采用溶栓前后质量减少计算溶栓率,统计学方法分析结果。结果光镜及扫描电镜下见各栓龄血栓溶栓后,血栓纤维网架被破坏。超声输出能量、微泡量、尿激酶浓度、溶栓时间4个因素对各栓龄血栓溶栓效果的影响均有统计学意义(P均<0.05);尿激酶对溶栓率的影响最大,最优溶栓参数组合为C4-D4-A1-B4,即:尿激酶800U/mL、溶栓时间40min、输出能量5%、微泡量400μL;4种因素各水平对4种栓龄血栓实验结果影响的效应曲线由高到低依次为3h、6h、12h、24h;不同栓龄血栓溶栓结果之间差异有统计学意义(P均<0.05)。结论诊断超声介导微泡造影剂具有助溶作用;在超声频率为1.82 MHz、能量输出为5%时,微泡已能被破坏而增强超声空化效应;固定超声频率,在高尿激酶浓度、延长溶栓时间、增大微泡量参数条件下可获得最优溶栓效果;随着血栓栓龄的增大,溶栓效果减低。Objective To investigate the efficacy of diagnostic ultrasound and microbubble contrast（MB） on enhancing thrombolysis in combination with urokinase（UK） and to determine the optimal combination for thrombolysis in vitro.Methods Four types of standardized red thrombus were prepared in vitro,including 3-hour-old（3 h）,6-hour-old（6 h）,12-hour-old（12 h）,and 24-hour-old（24 h）.The major parameters for the designed experiments included transmit powers of ultrasound（factor A,5%,25%,50%,100%）,MB volumes（factor B,50 μL,100 μL,200 μL,400 μL）,UK concentrations（factor C,100 U/mL,200 U/mL,400 U/mL,800 U/mL）,and lysis time（factor D,10 min,20 min,30 min,40 min）.An orthogonal array experimental design（OAD） based on four levels L16（45） of the above four parameters was employed to optimize the thrombolysis conditions.During the procedure of thrombolysis,the diagnostic ultrasound frequency was fixed at 1.82 MHz.The histopathological changes measured by HE staining and scanning electron microscope（SEM） were carried out to observe the clots before and after thrombolysis.The loss of clot weight before and after treatment was measured to determine the lysis efficiency（LE）.Analysis of variance（ANOVA） was performed to assess the LE according to the L16（45） matrix.Results The HE staining and SEM observation of thrombolysis under the following experimental conditions of 5% ultrasound transmit power,400 μL MB volume,800 U/mL UK concentration,and 40 min lysis time showed remarkable disaggregation of fibrin nets.The above four factors had significant impact on thrombus（all P0.05）, among which UK concentrations（factor C） w as the most significant one.The optimal scheme was determined as a C4-D4-A1B4 mode,with UK concentration 800 U/mL,lysis time 40 min,transmit power 5%,a nd MB volume 400 μL,respectively.The LE curves for 3h clots were superior to t he others.The lysis efficiencies for the clots showed significant differences a mong different type of thrombus（all P0.0

关 键 词：诊断超声 微泡造影剂 尿激酶 体外溶栓 栓龄 参数优化 

分 类 号：R445.1[医药卫生—影像医学与核医学]