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

发布时间：2018-04-02 12:38

本文选题：心肌梗塞　切入点：骨髓　出处：《中国组织工程研究》2017年17期

【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(P0.05);(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分数值有显著改善,差异有显著性意义(P0.05);(3)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平均毛细血管密度显著增多,差异有显著性意义(P0.05);(4)结果表明,超声介导一氧化氮微泡与冠脉移植骨髓间充质干细胞联合治疗可以改善家猪急性心肌梗死后的左心收缩功能,可能与超声介导一氧化氮微泡促进骨髓间充质干细胞向心肌缺血区归巢以及局部血管生成有关。
[Abstract]:Background: ultrasound combined with microbubble intervention has been found to enhance the efficacy of stem cell transplantation in the treatment of heart failure after myocardial infarction. However, it is not clear whether ultrasound mediated nitric oxide microbubble can be used in the treatment of myocardial infarction in large animals. Objective: to investigate the effect of ultrasound mediated oxidation of bone marrow mesenchymal stem cells (MSCs) after coronary artery transplantation in the treatment of myocardial infarction in large animals. The efficacy and possible mechanism of nitrogen microbubble and coronary artery transplantation bone marrow mesenchymal stem cells in the treatment of myocardial infarction in domestic pigs methods the density gradient centrifugation method was used to isolate the microbubbles and bone marrow mesenchymal stem cells. Culture, identification of bone marrow mesenchymal stem cells (CM-Dil) labeled bone marrow mesenchymal stem cells (BMSCs) in vitro, 24 swine myocardial infarction models were successfully established by interventional balloon occlusion of the anterior descending branch of the left coronary artery. 24 domestic pigs were randomly divided into PBS group. Bone marrow mesenchymal stem cells group, ultrasound mediated sonovir microbubble group, Ultrasound mediated nitric oxide (no) microbubble group was treated with the following intervention: 10 mL PBSs were injected into the coronary artery in the control group, and the mesenchymal stem cells group (1 脳 10 7) was diluted with 10 mL PBS and injected slowly through the coronary artery. The sonovir microbubbles were injected into the coronary artery in the sonovir microbubble group by coronary artery injection of 0.1 mL / kg, and then the bone marrow mesenchymal stem cells (about 1 脳 10 ~ (7)) were diluted with 10 mL PBS and slowly injected into the coronary artery. The methods and conditions of ultrasound mediated nitric oxide microbubble transplantation were the same as those of sonovir microbubble group. Only Sanovir microbubbles were changed into nitric oxide microbubbles. After 48 hours of intervention, three domestic pigs were randomly killed in each group, and the myocardial tissue of infarcted area was taken as frozen section. The left ventricular contractile function of each group was observed by M-mode echocardiography after 4 weeks. All the pigs were killed after cardiac function test. Myocardial tissue of infarcted area was stained with paraffin section and hematoxylin eosin staining. Results and conclusion the capillary density of myocardial ischemia area in each group was compared with that of sonovir microbubble group and bone marrow mesenchymal stem cell group, and compared with that of sonovir microbubble group and bone marrow mesenchymal stem cell group. The number of CM-Dil positive bone marrow mesenchymal stem cells (BMSCs) in ultrasound mediated nitric oxide microbubble group was significantly higher than that in sonovir microbubble group, bone marrow mesenchymal stem cell group and PBS group. The left ventricular ejection fraction (LVEF) was significantly improved in ultrasound mediated nitric oxide microbubble group (P 0.05), compared with sonovir microbubble group, bone marrow mesenchymal stem cell group and PBS group. The average capillary density in the nitric oxide microbubble group was significantly higher than that in the control group, and the difference was significant (P 0.05), the results showed that the average capillary density in the nitric oxide microbubble group was significantly higher than that in the control group. Ultrasound mediated nitric oxide microbubbles combined with bone marrow mesenchymal stem cells in coronary artery transplantation can improve left ventricular systolic function after acute myocardial infarction in domestic pigs. It may be related to ultrasound mediated nitric oxide microbubbles promoting homing of bone marrow mesenchymal stem cells to myocardial ischemic area and local angiogenesis.
【作者单位】：复旦大学附属上海市第五人民医院心内科;复旦大学附属中山医院心内科;东南大学附属中大医院心内科;南京大学医学院附属鼓楼医院急诊中心;
【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)~~
【分类号】：R542.22

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