超声针联合微泡干预尿激酶溶解体外血凝块

Ultrasound needle combined with microbubbles intervention of urokinase for lysis of in vitro blood clots

目的观察超声针联合微泡干预尿激酶溶解体外血凝块的效果。方法以10 ml牛全血制备体外血凝块,将40个血凝块随机分为5组(每组8个):对单纯超声组以超声针治疗8 min;对单纯尿激酶组注射2 ml尿激酶溶液(5000 IU/ml);对超声针+尿激酶组予以超声针治疗8 min+注射2 ml尿激酶溶液;对超声+生理盐水组予超声针治疗8 min+注射2 ml生理盐水;对超声针+尿激酶+微泡组于超声针+尿激酶组基础上向尿激酶溶液中加入0.01 ml自制微泡。以称重法计算各组血凝块溶解质量(W_(0))及溶解率;观察大体标本溶解情况,并于扫描电镜下观察血凝块微观变化。另取9个血凝块随机分为3组(每组3个):对照组,仅注入2 ml细胞膜特异性橙红色荧光染料1,1′-双十八烷基-3,3,3′,3′-四甲基吲哚羰花青高氯酸盐(DiI)溶液;超声组,超声针治疗8 min,同时注射2 ml DiI溶液;超声+微泡组,在超声组基础上于DiI溶液中额外加入0.01 ml自制微泡。于荧光显微镜下观察DiI渗透情况。结果干预前各组血凝块质量(W_(before))差异无统计学意义(P>0.05);干预后各组血凝块溶解质量(W_(0))及溶解率差异均有统计学意义(P均<0.01),超声+尿激酶组与超声+尿激酶+微泡组W_(0)及溶解率均显著高于其余3组(P均<0.05),但该2组间差异均无统计学意义(P均>0.05)。相比单纯超声组和单纯尿激酶组,超声+尿激酶组和超声+尿激酶+微泡组血凝块明显破坏,后者更为显著。超声组和超声+微泡组血凝块组织疏松,DiI渗透深远,满视野荧光,针道扩大,表面破坏,以超声+微泡组更显著。结论超声针能显著提高尿激酶对体外血凝块的溶解能力;联合应用微泡后溶解质量未见显著增加。

Objective To observe the intervention effect of ultrasound needle combined with microbubbles in urokinase for lysis of in vitro blood clots.Methods The in vitro blood clots were prepared with 10 ml bovine whole blood.Then 40 blood clots were randomly divided into 5 groups(each n=8),i.e.ultrasound alone group(only ultrasound needle for 8 min),urokinase alone group(injection of 2 ml urokinase solution[5000 IU/ml]via needle sheath),ultrasound+urokinase group(8 min ultrasound needle and injection of 2 ml urokinase solution),ultrasound+normal saline group(8 min ultrasound needle and injection of 2 ml normal saline)and ultrasound+urokinase+microbubbles group(0.01 ml self-made microbubbles added into urokinase solution on the basis of ultrasound needle+urokinase group).The lysis weight(W_(0))and lysis rate were calculated with weighing method.Then macroscopic lysis performances of gross specimens were evaluated,and the samples were prepared for observing microscopic changes under scanning electron microscopy.Other clots were reprepared and divided into 3 groups(each n=3),i.e.control group(injection of 2 ml of membrane-specific orange-red fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate[DiI]),ultrasound group(8 min ultrasound needle and injection of 2 ml DiI solution)and ultrasound+microbubbles group(added 0.01 ml self-made microbubbles into DiI solution on the basis of ultrasound needle+urokinase group).After sections,the penetration result of fluorescent dye DiI was assessed under microscope.Results Clot weights before intervention(W_(before))were not significantly different among 5 groups(P>0.05),while W_(0) and lysis rates were significantly different among groups after intervention(both P<0.01).Both W_(0) and lysis rate in ultrasound+urokinase group and ultrasound+urokinase+microbubbles group were higher than those in the other groups(all P<0.05),while no statistical difference was found between ultrasound+urokinase group and ultrasound+urokinase+microbubbles group(both P>0.05).The destruction of clots in ultrasound+urokinase group and ultrasound+urokinase+microbubbles group were stronger than in ultrasound alone group and urokinase alone group,and in ultrasound+urokinase+microbubbles group was more significantly obvious.Loose tissue were found in both ultrasound group and ultrasound+microbubbles group,also enhanced DiI penetration with full field fluorescence,and enlarged needle path with apparent surface damage,especial in the latter.Conclusion Ultrasound needle could significantly improve urokinase for thrombolysis of in vitro clots,but the addition of microbubbles did not greatly increase the lysis weight.