诊断超声联合微泡对骨髓间充质干细胞移植改善兔心肌缺血灌注的研究

Mesenchymal stem cells transplantation under the diagnostic ultrasound mediated microbubbles destruction in improving myocardial perfusion after rabbit myocardial ischemia

目的探讨诊断超声联合微泡介导下经静脉移植同种异体骨髓间充质干细胞（MSCs）改善心肌缺血兔血流灌注的价值。方法冠状动脉左前降支结扎法建立兔心肌缺血模型，分为对照组（组Ⅰ）、单纯静脉移植MSCs组（组Ⅱ）和超声＋微泡＋MSCs组（组Ⅲ）。4周后行心肌超声造影（MCE）分析各组心肌缺血区血流灌注情况，超声心动图检测左心室功能，免疫组织化学检测心肌缺血区CD34表达，Western blotting检测VEGF蛋白的表达并行定量分析。结果MCE检测前壁的平均灰阶值组I（51．86±11．75）与组Ⅱ（63．08±15．32）和组Ⅲ（70．42±12．21）比较差异有统计学意义（P值分别为0．032，0．000）。左室短轴缩短率（FS）组Ⅰ[（19．28±2．84）％]与组Ⅱ[（26．29±2．93）％]和组Ⅲ[（30．43±4．09）％]比较差异均有统计学意义（P〈0．01，P〈0．05），组Ⅱ和组Ⅲ比较差异亦有统计学意义（P〈0．05）。组Ⅲ的射血分数（EF）为（61．5±5．8）％，与组I[（42．6±5．0）％]和组Ⅱ[（55．6±4．71）％]比较差异均有统计学意义（P＜0．01，P＜0．05）。双平面Simpson法EF值组ⅢU48．6±3．96）％]与组Ⅰ[（34．64±4．59）％]和组Ⅱ[41．78±4．21）％]比较差异有统计学意义（P＜0．01，P＜0．05）。CD34免疫组织化学检测显示各组均可见阳性血管着色，但组Ⅲ阳性血管最多，组Ⅱ次之，组I最少。WesternblottingVEGF／GAPDH定量分析显示，组Ⅲ表达（236．59±47．13）明显高于组Ⅱ（151．48±25．07，P＜0．01）和组I（89．43±20．43，P〈0．01）。结论超声介导的微泡破坏与经静脉移植MSCs可有效改善心肌缺血兔的血流灌注及心功能。

Objective To explore the value of diagnostic ultrasound mediated microbubble destruction in improving .the myocardial perfusion and left ventricular systolic function when cooperated with the mecsenchymal stem cells（MSCs） transplantation in rabbit myocardial isehemia. Methods One week after myocardial ischemia （MI） modeling,36 rabbits were divided into 3 groups,the control group（group Ⅰ） ,intravenous injection of MSCs group（group Ⅱ ） and ultrasound ＋ microbubble ＋ MSCs group （group Ⅲ）. Myocardial contrast enhancement （MCE） was performed and quantification analysis of anterior wall was assessed with Photoshop. Left ventricular systolic function was assessed with M-mode echocardiography and bi-plane Simpson＇s method. CD34 expression in heart was detected with immunohistochemisty（IHC）. Western blotting was applied to detect the level of VEGF in three groups. Results The differences of gray scale analyzed with histogram of Photoshop in anterior wall of ischemia myocardium between the group Ⅰ and group Ⅱ or group Ⅲ were significant,and P value was 0. 032 and 0. 000,respectively. There were significant differences of FS between group Ⅲ （30.43 ±4.09）% and group Ⅱ（26. 29 ±2.93） %, p 〈0. 01 ,and similar to group I （19.28 ± 2. 84） ~. The difference of EF（%） between group Ⅲ and group Ⅱwas significant E（61.5± 5.8 vs 53.6 ±4.71）, P 〈0.05%,or markedly significant between group Ⅲ and group I E（61.5 ± 5.8 vs 42.6 ± 5.0）, P d0. 011. EF（%） assessed with bi-plane Simpson＇s method was significantly increased from （34.64 ± 4.59） in group Ⅰ to （41.78 ± 4.21） in group Ⅱ and （48.6 ± 3.96） in groupⅢ. The expression of CD34 assessed with immunohistochemistry was the highest in groupⅢ . The level of VEGF with western blotting in group Ⅲ was significantly higher than other two groups. Conclusions It is an efficacious transplantation means of MSCs infusion under the ultrasound mediated microbubles destruction in improving the myocardial perfusion and cardiac systolic function.