超声介导一氧化氮微泡辅助骨髓间充质干细胞移植治疗心肌梗死  

Ultrasound mediated nitric oxide microbubbles enhance the therapeutic efficacy of bone marrow mesenchymal stem cell transplantation on myocardial infarctions

作　　者：陈飞[1] 许澎[1] 乔祺[1] 樊冰[2] 童嘉毅[3] 范国峰[4] Chen Fei Xu Peng Qiao Qi Fan Bing Tong Jia-yi Fan Guo-feng(Department of Cardiology, Fifth People＇s Hospital of Fudan University, Shanghai 200240, China Department of Cardiology, Zhongshan Hospital of Fudan University, Shanghai 200032, China Department of Cardiology, Zhongda Hospital of Southeast University, Nanjing 210007, Jiangsu Province, China Emergency Center, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China)

机构地区：[1]复旦大学附属上海市第五人民医院心内科,上海市200240 [2]复旦大学附属中山医院心内科,上海市200032 [3]东南大学附属中大医院心内科,江苏省南京市210007 [4]南京大学医学院附属鼓楼医院急诊中心,江苏省南京市210008

出　　处：《中国组织工程研究》2017年第17期2678-2683,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81070265/H0222);江苏省卫生厅青年科研课题(Q201306);南京市医学科技发展基金(YKK16100);上海市闵行区自然科学基金(2014MHZ037)~~

摘　　要：背景:多项实验研究发现,超声联合微泡干预能够增强干细胞移植治疗心肌梗死后心力衰竭的效果,具有良好的应用前景。但超声介导一氧化氮微泡经冠脉移植骨髓间充质干细胞治疗大型动物心肌梗死是否具有同样的效果仍不明确。目的:探讨超声介导一氧化氮微泡与冠脉移植骨髓间充质干细胞治疗家猪心肌梗死的有效性及可能机制。方法:(1)密度梯度离心法分离、培养、鉴定家猪骨髓间充质干细胞,CM-Dil体外标记骨髓间充质干细胞;(2)经介入法球囊封堵猪左冠状动脉前降支成功建立24只家猪心肌梗死模型,随机分为PBS组、骨髓间充质干细胞组、超声介导声诺维微泡组、超声介导一氧化氮微泡组(n=6)。于造模后1周进行如下干预:PBS组经冠脉注射10 m L PBS,骨髓间充质干细胞组将骨髓间充质干细胞(约1×10~7)用10 m L PBS稀释后经冠脉缓慢注射;超声介导声诺维微泡组经冠脉注入声诺维微泡0.1 m L/kg,同时超声干预(1 MHz、2 W/cm^2)2 min,然后将骨髓间充质干细胞(约1×10~7)用10 m L PBS稀释后经冠脉缓慢注射;超声介导一氧化氮微泡组移植方法及条件同超声介导声诺维微泡组,仅将声诺维微泡换作一氧化氮微泡;(3)干预后48 h随机处死每组各3只家猪,取梗死区心肌组织作冰冻切片,计数比较各组缺血心肌内荧光标记阳性骨髓间充质干细胞。4周后对每组剩余家猪行M型心脏超声观察比较各组左心室收缩功能。心功能检测结束后处死所有家猪,取梗死区心肌组织行石蜡切片及苏木精-伊红染色,计数比较各组心肌缺血区的毛细血管密度。结果与结论:(1)与超声介导声诺维微泡组及骨髓间充质干细胞组比较,超声介导一氧化氮微泡组CM-Dil阳性骨髓间充质干细胞数增多,差异有显著性意义(P<0.05);(2)与超声介导声诺维微泡组、骨髓间充质干细胞组及PBS组比较,超声介导一氧化氮微泡组左室射血�BACKGROUND： Recent experimental studies have found ultrasound mediated microbubbles potentiate stem cell therapy in myocardial infarction （MI）-induced heart failure, indicating a good application prospect. But whether ultrasound mediated nitric oxide （NO） microbubbles also have the same effect in the intracoronary transplantation of bone marrow mesenchymal stem cells （BMSCs） for treatment of large animals with MI is still unknown. OBJECTIVE： To investigate the effectiveness and possible mechanism of ultrasound mediated NO microbubbles in potentiating intracoronally transplanted BMSCs homing to the infarcted area in a MI pig model.METHODS： Density gradient centrifugation culture method was used in the isolation and cultivation of BMSCs. CM-Dil was used to label BMSCs in vitro. Twenty-four pigs were used to make MI models by blocking the left anterior descending coronary artery, and then were divided into PBS group, BMSCs group, ultrasound＋microbubbles＋BMSCs（MB） group, ultrasound＋NO microbubbles＋BMSCs （NO-MB） group（n=6 per group）. In the PBS group, 10 mL of PBS was intracoronally injected. In the BMSCs group, about 1×107 BMSCs were diluted in 10 mL of PBS and then intracoronally infused. In the MB group, 0.1 mL/kg sulphur hexafluoride microbubbles （Sono Vue） was intracoronally injected together with ultrasound treatment （1 MHz, 2 W/cm2, 2 minutes）, followed by intracoronary infusion of about 1×107 BMSCs that were diluted in 10 mL of PBS. In the NO-MB group, all methods and conditions were identical to those in the MB group except only 0.1 mL/kg of Sono Vue was replaced by 0.1 mL/kg NO microbubbles. Three pigs were sacrificed in each group 48 hours after CM-Dil positive BMSCs transplantation. The labeled BMSCs were observed and counted by fluorescent microscope after frozen sectioning of the infarct area. We assessed and compared left ventricular systolic function with M-mode ultrasound among groups at 4 weeks after intervention. After cardiac function test, the rest p

关 键 词：干细胞 移植 骨髓间充质干细胞 心肌梗死 超声 一氧化氮微泡 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]