本文選題：心肌梗塞　切入點(diǎn)：骨髓　出處：《中國組織工程研究》2017年17期

【摘要】：背景:多項(xiàng)實(shí)驗(yàn)研究發(fā)現(xiàn),超聲聯(lián)合微泡干預(yù)能夠增強(qiáng)干細(xì)胞移植治療心肌梗死后心力衰竭的效果,具有良好的應(yīng)用前景。但超聲介導(dǎo)一氧化氮微泡經(jīng)冠脈移植骨髓間充質(zhì)干細(xì)胞治療大型動(dòng)物心肌梗死是否具有同樣的效果仍不明確。目的:探討超聲介導(dǎo)一氧化氮微泡與冠脈移植骨髓間充質(zhì)干細(xì)胞治療家豬心肌梗死的有效性及可能機(jī)制。方法:(1)密度梯度離心法分離、培養(yǎng)、鑒定家豬骨髓間充質(zhì)干細(xì)胞,CM-Dil體外標(biāo)記骨髓間充質(zhì)干細(xì)胞;(2)經(jīng)介入法球囊封堵豬左冠狀動(dòng)脈前降支成功建立24只家豬心肌梗死模型,隨機(jī)分為PBS組、骨髓間充質(zhì)干細(xì)胞組、超聲介導(dǎo)聲諾維微泡組、超聲介導(dǎo)一氧化氮微泡組(n=6)。于造模后1周進(jìn)行如下干預(yù):PBS組經(jīng)冠脈注射10 m L PBS,骨髓間充質(zhì)干細(xì)胞組將骨髓間充質(zhì)干細(xì)胞(約1×10~7)用10 m L PBS稀釋后經(jīng)冠脈緩慢注射;超聲介導(dǎo)聲諾維微泡組經(jīng)冠脈注入聲諾維微泡0.1 m L/kg,同時(shí)超聲干預(yù)(1 MHz、2 W/cm~2)2 min,然后將骨髓間充質(zhì)干細(xì)胞(約1×10~7)用10 m L PBS稀釋后經(jīng)冠脈緩慢注射;超聲介導(dǎo)一氧化氮微泡組移植方法及條件同超聲介導(dǎo)聲諾維微泡組,僅將聲諾維微泡換作一氧化氮微泡;(3)干預(yù)后48 h隨機(jī)處死每組各3只家豬,取梗死區(qū)心肌組織作冰凍切片,計(jì)數(shù)比較各組缺血心肌內(nèi)熒光標(biāo)記陽性骨髓間充質(zhì)干細(xì)胞。4周后對每組剩余家豬行M型心臟超聲觀察比較各組左心室收縮功能。心功能檢測結(jié)束后處死所有家豬,取梗死區(qū)心肌組織行石蠟切片及蘇木精-伊紅染色,計(jì)數(shù)比較各組心肌缺血區(qū)的毛細(xì)血管密度。結(jié)果與結(jié)論:(1)與超聲介導(dǎo)聲諾維微泡組及骨髓間充質(zhì)干細(xì)胞組比較,超聲介導(dǎo)一氧化氮微泡組CM-Dil陽性骨髓間充質(zhì)干細(xì)胞數(shù)增多,差異有顯著性意義(P0.05);(2)與超聲介導(dǎo)聲諾維微泡組、骨髓間充質(zhì)干細(xì)胞組及PBS組比較,超聲介導(dǎo)一氧化氮微泡組左室射血分?jǐn)?shù)值有顯著改善,差異有顯著性意義(P0.05);(3)與超聲介導(dǎo)聲諾維微泡組、骨髓間充質(zhì)干細(xì)胞組及PBS組比較,超聲介導(dǎo)一氧化氮微泡組平均毛細(xì)血管密度顯著增多,差異有顯著性意義(P0.05);(4)結(jié)果表明,超聲介導(dǎo)一氧化氮微泡與冠脈移植骨髓間充質(zhì)干細(xì)胞聯(lián)合治療可以改善家豬急性心肌梗死后的左心收縮功能,可能與超聲介導(dǎo)一氧化氮微泡促進(jìn)骨髓間充質(zhì)干細(xì)胞向心肌缺血區(qū)歸巢以及局部血管生成有關(guān)。
[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.
【作者單位】： 復(fù)旦大學(xué)附屬上海市第五人民醫(yī)院心內(nèi)科;復(fù)旦大學(xué)附屬中山醫(yī)院心內(nèi)科;東南大學(xué)附屬中大醫(yī)院心內(nèi)科;南京大學(xué)醫(yī)學(xué)院附屬鼓樓醫(yī)院急診中心;
【基金】：國家自然科學(xué)基金(81070265/H0222) 江蘇省衛(wèi)生廳青年科研課題(Q201306) 南京市醫(yī)學(xué)科技發(fā)展基金(YKK16100) 上海市閔行區(qū)自然科學(xué)基金(2014MHZ037)~~
【分類號】：R542.22

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