Overexpression of microRNA-124 promotes the neuronal differentiation of bone marrow-derived mesenchymal stem cells

microRNAs （miRNAs） play an important regulatory role in the self-renewal and differentiation of stem cells. In this study, we examined the effects of miRNA-124 （miR-124） overexpression in bone marrow-derived mesenchymal stem cells. In particular, we focused on the effect of overexpression on the differentiation of bone marrow-derived mesenchymal stem cells into neurons. First, we used GeneChip technology to analyze the expression of miRNAs in bone marrow-derived mesen- chymal stem cells, neural stem cells and neurons, miR-124 expression was substantially reduced in bone marrow-derived mesenchymal stem cells compared with the other cell types. We con- structed a lentiviral vector overexpressing miR-124 and transfected it into bone marrow-derived mesenchymal stem cells. Intracellular expression levels of the neuronal early markers [3-III tu- bulin and microtubule-associated protein-2 were significantly increased, and apoptosis induced by oxygen and glucose deprivation was reduced in transfected cells. After miR-124-transfected bone marrow-derived mesenchymal stem cells were transplanted into the injured rat spinal cord, a large number of cells positive for the neuronal marker neurofilament-200 were observed in the transplanted region. The Basso-Beattie-Bresnahan locomotion scores showed that the motor function of the hind limb of rats with spinal cord injury was substantially improved. These re- sults suggest that miR-124 plays an important role in the differentiation of bone marrow-derived mesenchymal stem cells into neurons. Our findings should facilitate the development of novel strategies for enhancing the therapeutic efficacy of bone marrow-derived mesenchymal stem cell transplantation for spinal cord injury.

Transplantation of placenta-derived mesenchymal stem cell-induced neural stem cells to treat spinal cord injury

Because of their strong proliferative capacity and multi-potency, placenta-derived mesenchymal stem cells have gained interest as a cell source in the field of nerve damage repair. In the present study, human placenta-derived mesenchymal stem ceils were induced to differentiate into neural stem cells, which were then transplanted into the spinal cord after local spinal cord injury in rats. The motor functional recovery and pathological changes in the injured spinal cord were observed for 3 successive weeks. The results showed that human placenta-derived mesenchymal stem cells can differentiate into neuron-like cells and that induced neural stem cells contribute to the restoration of injured spinal cord without causing transplant rejection. Thus, these cells promote the recovery of motor and sensory functions in a rat model of spinal cord injury. Therefore, human placenta-derived mesenchymal stem cells may be useful as seed cells during the repair of spinal cord injury.

Nutritional assessment with different tools in leukemia patients after hematopoietic stem cell transplantation

Objective： Correct nutritional assessment is essential for leukemia patients after hematopoietic stem cell transplantation （HSCT）. This study aimed to investigate the best nutritional assessment method for leukemia patients after HSCT, and find the possible nutritional risk of the patients during the transplantation process in order to intervene in the patients with nutritional risks and undernourished patients timely, so that the entire transplantation process could be successfully completed. Methods： A prospective study was performed in 108 leukemia patients after HSCT, and different nutritional assessment methods, including nutritional risk screening 2002 （NRS2002）, mini nutritional assessment （MNA）, subjective globe assessment （SGA） and malnutritional universal screening tools （MUST）, were used. The associations between nutritional status of these patients and nutritional assessment methods were analyzed. Results： A total of 108 patients completed SGA, and 99 patients completed NRS2002, MNA and MUST. During the treatment process, 85.2% of the patients lost weight, wherein, 50% lost weight greater than 5%, and 42.6% had significantly reduced food intake. For nutritional risk assessment, the positive rates of NRS2002, MNA and MUST were 100%, 74.7% and 63.6%, respectively. There was a significant difference （P〈0.05） among the positive rates of NRS2002, MNA and MUST. In undernutrition assessment, the positive rate of SGA （83.3%） was significantly higher than that of MNA （17.2%） （P〈0.05）, and the incidence rate of nutritional risk among leukemia patients _〈30 years old was greater than that of patients 〉30 years old （P〈0.05）. Conclusions： Patients with leukemia were in poor nutritional status during and after HSCT. The leukemia patients 〈30 years old had a greater incidence rate of nutritional risk. As nutritional risk screening tool, the specificity of NRS2002 is not high, but it can be used for evaluating nutritional deficiencies. MNA is a good nutritional risk screening tool, but not an adequate tool for nutritional assessment. If assessment of undernutrition is necessary, the combination of all these screening tools and clinical laboratory indicators should he applied to improve accuracy.

Are human dental papilla-derived stem cell and human brain-derived neural stem cell transplantations suitable for treatment of Parkinson’s disease?

Transplantation of neural stem cells has been reported as a possible approach for replacing impaired dopaminergic neurons. In this study, we tested the efficacy of early-stage human dental papilla-derived stem cells and human brain-derived neural stem cells in rat models of 6-hydroxydopamine-induced Parkinson＇s disease. Rats received a unilateral injection of 6-hydroxydopamine into right medial forebrain bundle, followed 3 weeks later by injections of PBS, early-stage human dental papilla-derived stem cells, or human brain-derived neural stem cells into the ipsilateral striatum. All of the rats in the human dental papilla-derived stem cell group died from tumor formation at around 2 weeks following cell transplantation. Postmortem examinations revealed homogeneous malignant tumors in the striatum of the human dental papilla-derived stem cell group. Stepping tests revealed that human brain-derived neural stem cell transplantation did not improve motor dysfunction. In apomorphine-induced rotation tests, neither the human brain-derived neural stem cell group nor the control groups （PBS injection） demonstrated significant changes. Glucose metabolism in the lesioned side of striatum was reduced by human brain-derived neural stem cell transplantation. [18F]-FP-CIT PET scans in the striatum did not demonstrate a significant increase in the human brain-derived neural stem cell group. Tyrosine hydroxylase （dopaminergic neuronal marker） staining and G protein-activated inward rectifier potassium channel 2 （A9 dopaminergic neuronal marker） were positive in the lesioned side of striatum in the human brain-derived neural stem cell group. The use of early-stage human dental papilla-derived stern cells confirmed its tendency to form tumors. Human brain-derived neural stem cells could be partially differentiated into dopaminergic neurons, but they did not secrete dopamine.

In vivo tracking of neuronal-like cells by magnetic resonance in rabbit models of spinal cord injury

In vitro experiments have demonstrated that neuronal-like cells derived from bone marrow mesen- chymal stem cells can survive, migrate, integrate and help to restore the function and behaviors of spinal cord injury models, and that they may serve as a suitable approach to treating spinal cord injury. However, it is very difficult to track transplanted cells in vivo. In this study, we injected su- perparamagnetic iron oxide-labeled neuronal-like cells into the subarachnoid space in a rabbit model of spinal cord injury. At 7 days after cell transplantation, a small number of dot-shaped low signal intensity shadows were observed in the spinal cord injury region, and at 14 days, the number of these shadows increased on T2-weighted imaging. Perl＇s Prussian blue staining detected dot-shaped low signal intensity shadows in the spinal cord injury region, indicative of superpara- magnetic iron oxide nanoparticle-labeled cells. These findings suggest that transplanted neu- ronal-like cells derived from bone marrow mesenchymal stem cells can migrate to the spinal cord injury region and can be tracked by magnetic resonance in vivo. Magnetic resonance imaging represents an efficient noninvasive technique for visually tracking transplanted cells in vivo.

Effect of VEGF on Neural Differentiation of Human Embryonic Stem Cells in vitro

The effects of vascular endothelial growth factor （VEGF） on neural differentiation of human embryonic stem cells （hESCs） in vitro and the possible mechanism were observed. The hESCs lines, TJMU1 and TJMU2, were established and stored by our laboratory, hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups： group A, routine induction; group B, routine induction＋10 ng/mL VEGF; group C, routine in- duction＋10 ng/mL VEGF＋10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were de- tected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence. The percentage of Nestin positive cells in group B was significantly higher than in groups A and C, while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C （P〈0.01）. There was no significant difference between groups A and C （P〉0.05）. It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

Similarity on neural stem cells and brain tumor stem cells in transgenic brain tumor mouse models

Although it is believed that glioma is derived from brain tumor stem cells, the source and molecular signal pathways of these cells are still unclear. In this study, we used stable doxycycline-inducible transgenic mouse brain tumor models （c-myc/SV40Tag＋/Tet-on＋） to explore the malignant trans- formation potential of neural stem cells by observing the differences of neural stem cells and brain tumor stem cells in the tumor models. Results showed that chromosome instability occurred in brain tumor stem cells. The numbers of cytolysosomes and autophagosomes in brain tumor stem cells and induced neural stem cells were lower and the proliferative activity was obviously stronger than that in normal neural stem cells. Normal neural stem cells could differentiate into glial fibrillary acidic protein-positive and microtubule associated protein-2-positive cells, which were also negative for nestin. However, glial fibrillary acidic protein/nestin, microtubule associated protein-2/nestin, and glial fibrillary acidic protein/microtubule associated protein-2 double-positive cells were found in induced neural stem cells and brain tumor stem cells. Results indicate that induced neural stem cells are similar to brain tumor stem cells, and are possibly the source of brain tumor stem cells.

A brief review of recent advances in stem cell biology

Stem cells have the remarkable potential to develop into many different cell types, essentially with- out limit to replenish other cells as long as the person or animal is still alive, offering immense hope of curing Alzheimer＇s disease, repairing damaged spinal cords, treating kidney, liver and lung diseases and making damaged hearts whole. Until recently, scientists primarily worked with two kinds of stem cells from animals and humans： embryonic stem cells and non-embryonic ＂somatic＂ or ＂adult＂ stem cells. Recent breakthrough make it possible to convert or ＂reprogram＂ specialized adult cells to assume a stem stem-like cells with different technologies. The review will briefly dis- cuss the recent progresses in this area.

体外诱导大鼠骨髓间质干细胞分化为胰岛素分泌细胞(英文)

目的探索大鼠骨髓间质干细胞体外诱导分化为胰岛素分泌细胞的方法。为解决胰岛移植物来源匮乏这一问题提供新的思路。方法 采用横向分化技术,将成年大鼠骨髓间质干细胞诱导成为胰岛素分泌细胞。间接免疫荧光法鉴定诱导前后细胞nestin、胰岛素、胰高血糖素、生长抑素及Pdx-1的表达,RT-PCR法检测诱导前后细胞nestin,胰岛素-1,葡萄糖转运子-2,葡萄糖激酶及其转录因子Isl-1,Pdx-1,Pax-4和Pax-6 mRNA的表达;测定24 h胰岛素分泌量和胰岛素刺激实验评价诱导前后细胞的功能。结果 诱导5 h,nestin阳性细胞为(44.6±7.3)％;诱导24 h,nestin阳性细胞增至(61.8±8.4)％;此后,nestin阳性细胞数目开始下降,诱导第14天后,nestin表达基本消失。诱导后的胰岛素分泌细胞可以表达胰岛素、胰高血糖素、生长抑素和Pdx-1等蛋白;表达胰岛素-1、葡萄糖转运子-2、葡萄糖激酶及其多种转录因子mRNA;胰岛素分泌量增加;胰岛素刺激实验反应敏感等。而诱导前MSCc不县备上述特点。结论 大鼠骨髓间质干细胞体外可以诱导成为胰岛素分泌细胞,为胰岛移植开辟新的研究思路。

转染IL-12基因的人脐带间充质干细胞对卵巢癌细胞的体外增殖和对裸鼠体内移植瘤的抑制作用

目的探讨人脐带间充质干细胞(UC-MSCs)负载白细胞介素12(IL-12)重组腺病毒载体抗卵巢癌SKOV3作用的研究。方法腺病毒介导IL-12基因转染体外培养的UC-MSCs(AdIL-12-MSCs),Western blotting和RT-PCR检测AdIL-12-MSCs中IL-12基因蛋白及mRNA表达,ELISA法检测AdIL-12-MSCs上清液中IL-12的表达。光镜下观察外源性IL-12对SKOV3细胞形态的影响,MTT法检测AdIL-12-MSCs分泌的IL-12对SKOV3细胞增殖活性的影响,流式细胞仪检测AdIL-12-MSCs上清液对SKOV3细胞凋亡的影响。建立裸鼠卵巢癌SKOV3皮下移植瘤模型,观察移植AdIL-12-MSCs后对移植瘤的影响。结果腺病毒介导IL-12基因成功转染UC-MSCs形成AdIL-12-MSCs,转染后细胞IL-12基因在蛋白及mRNA水平均有明显表达。AdIL-12-MSCs分泌的外源性IL-12显著抑制卵巢癌SKOV3细胞的增殖并诱导其凋亡,并抑制卵巢癌SKOV3移植瘤在裸鼠体内的生长(P<0.05)。结论转染IL-12基因的UC-MSCs能够在蛋白及mRNA水平表达IL-12基因,显著抑制卵巢癌SKOV3细胞的增殖并诱导其凋亡,抑制卵巢癌SKOV3移植瘤在裸鼠体内的生长。

翼状胬肉手术治疗研究进展

翼状胬肉是一种仅见于人类的常见的眼表疾病之一。其发病机制有多种不同的解释,临床治疗效果不尽人意,复发率较高。几十年来国内外学者对该疾病的手术治疗方法做了大量的研究,以期达到更小的手术损伤和更低的复发率。传统手术方法损伤大,复发率较高。激光治疗具有手术损伤小、安全性高的优点,但其远期疗效,复发率及与其他方法如丝裂霉素,羊膜等联合治疗的效果有待观察。

兔脂肪干细胞的分离培养鉴定及成骨诱导分化研究

[目的]探讨在成骨诱导条件下兔脂肪干细胞的体外诱导分化情况。[方法]取3个月龄日本大耳白兔颈背部皮下脂肪,用Ⅰ型胶原酶消化获得细胞。Stro-1免疫细胞化学染色鉴定细胞性质;加入成骨诱导液后依次进行形态学、Ⅰ型胶原、碱性磷酸酶及钙盐沉积相关检测。[结果](1)原代所获细胞Stro-1表达阳性。(2)在诱导条件下,Ⅰ型胶原、碱性磷酸酶及钙盐沉积均呈阳性表达。[结论]脂肪干细胞来源广、易获取、对机体创伤小,与骨髓间充质干细胞类似,经体外诱导后可实现成骨分化,为骨组织工程提供了一种新的种子细胞。

亲缘与非亲缘供者造血干细胞动员和采集的安全性比较

本研究对捐献骨髓及外周造血干细胞的健康亲缘供者及只捐献外周造血干细胞的非亲缘供者,在造血干细胞动员和采集的安全性方面进行比较。对2005年9月至2006年8月在北京大学人民医院血液病研究所提供异基因造血干细胞的亲缘供者100例及2003年11月至2007年12月在中国造血干细胞捐献者资料库北京管理中心登记的非血缘供者71例,在造血干细胞动员、采集及采集后1、3、6个月及每年进行了评估。对血常规指标、不良反应等进行观察记录,并对随访期间的长期不良反应及生活质量进行了问卷调查。结果显示:亲缘供者提供的骨髓+外周血干细胞总MNC剂量为6.70(4.11-12.23)×108/kg,总CD34+细胞剂量为3.40(1.61-13.57)×106/kg;非亲缘供者提供的外周血干细胞总MNC剂量为6.69(3.35-11.48)×108/kg,总CD34+细胞剂量为3.50(1.15-11.60)×106/kg。动员时的常见副作用为骨痛,在亲缘供者的发生率为47%,在非亲缘供者的发生率为43.7%,两组之间无显著性差异;采集时的常见副作用为感觉异常(口唇和四肢),在亲缘供者的发生率为25%,在非亲缘供者的发生率为29.6%,两组之间无显著性差异;所有供者对副作用皆可耐受,没有供者因为不能耐受而中断采集。亲缘供者由于骨髓和外周血的采集,其血红蛋白水平低于非亲缘供者[(125.8±20.2)g/Lvs(143.2±20.1)g/L](p<0.05)。非亲缘供者由于外周干细胞采集多为2次,其血小板计数低于亲缘供者[(126.2±57.2)×109/Lvs(162.4±72.9)×109/L](p<0.05)。在长期随访中,亲缘供者与非亲缘供者的血常规检查结果比较无显著性差异,无长期的不良反应,健康状况良好。结论:亲缘与非亲缘供者进行造血干细胞采集都是安全可行的。术前进行完备的检查,术中仔细操作、严密观察,及术后长期随访对于供者的安全有重要的意义。

神经干细胞移植脑出血恢复期大鼠神经功能和促血管生成素1及受体的表达

背景:近年研究发现骨髓间充质干细胞经过长时间体外培养后,可自然分化为神经干细胞,从而再定向分化为神经细胞及神经胶质细胞,为帕金森病、脑梗死后遗症、小脑萎缩和脑发育不良等疾病的治疗提供了一种新的思路。目的:探讨神经干细胞移植对脑出血恢复期大鼠神经功能的影响及其作用机制。方法:将60只雄性SD大鼠按照随机数字表法分为正常组(18只)、脑出血组(21只)和移植组(21只),后2组大鼠采用Ⅶ型胶原酶诱导法建立大鼠脑出血模型,造模21 d后移植组大鼠通过尾静脉注射神经干细胞,脑出血组给予等量生理盐水。移植后第7,14,21天进行改良黏附物移除试验(MST)评分,评分结束后处死各组大鼠,采用RT-PCR法测定大鼠出血周围脑组织促血管生成素1 mR NA表达,采用Western Blotting法检测大鼠出血周围脑组织酪氨酸激酶受体2蛋白表达。结果与结论:与正常组相比,脑出血组和移植组各时点MST评分均明显降低,差异均有显著性意义(P<0.05);从移植后7 d开始移植组各时点的MST评分均明显高于脑出血组,差异均有显著性意义(P<0.05)。移植后7,14,21 d移植组和脑出血组促血管生成素1 m RNA和酪氨酸激酶受体2蛋白含量均明显高于正常组,且移植组升高更为明显,差异均有显著性意义(P均<0.05)。结果表明神经干细胞移植能够有效促进脑出血恢复期大鼠神经功能的恢复,其机制可能与增加出血周围脑组织促血管生成素及酪氨酸激酶受体2的含量有关。

三氧化二砷联合FLAG方案治疗复发急性髓性白血病及对白血病干细胞的影响

目的:探讨三氧化二砷(Arsenic Trioxide,AT)联合改良氟达拉滨+阿糖胞苷+粒系集落刺激因子(FLAG)方案治疗复发难治急性髓性白血病(AML)的疗效以及对白血病干细胞(leukemia stem cells,LSC)的作用。方法:10例难治和复发AML患者采用AT联合FLAG方案治疗,观察临床治疗效果,并检测治疗前后患者骨髓中LSC和P-gp阳性细胞数的变化。结果:10例患者经AT联合FLAG方案治疗,持续缓解时间平均8个月(4～12个月),完全缓解率(CR)3个月、6个月和12个月分别为80%、60%和25%。10例患者均出现了Ⅳ度骨髓抑制,但未出现其它严重不良反应,无1例在治疗过程中死亡。骨髓中P-gp^+细胞由治疗前的(23.55±2.75)%降为治疗后的(11.67±3.50)%;骨髓中LSC(CD34^+CD38^-CD123^+)的相对含量显著降低,治疗前和治疗后分别为(4.30±1.30)%和(2.60±0.70)%。结论:AT联合改良FLAG方案可有效诱导难治和复发急性髓性白血病患者缓解,并降低患者骨髓中白血病干细胞(LSC)和P-gp^+细胞的数量。

淫羊藿总黄酮对衰老大鼠小肠干细胞及其微环境的调节作用

目的:探讨淫羊藿总黄酮对衰老大鼠小肠干细胞及其微环境变化的调节作用。方法:SD大鼠按其不同发育阶段分为6月龄成年组、24月龄衰老组及淫羊藿总黄酮低、中、高剂量组。采用常规HE染色,高碘酸-Schiff试剂(PAS)染色,免疫组化(荧光),Western bolt检测各组别小肠干细胞数量,增殖分化功能及其微环境相关信号的变化。结果:与成年组比较,衰老组小肠绒毛长度、隐窝深度、V/C值、杯状细胞数量及PCNA表达量降低,小肠干细胞数量显著减少,干细胞微环境Wnt/β-catenin和BMP信号均呈下调趋势。给予淫羊藿总黄酮后,与衰老组比较,小肠隐窝干细胞数量有所增加,增殖分化功能有所提升,表现为绒毛长度、隐窝深度、V/C值、杯状细胞数量及PCNA表达量增加,小肠干细胞微环境Wnt/β-catenin和BMP信号均呈上调趋势。结论:衰老过程中,小肠干细胞数量减少,增殖分化功能降低,干细胞微环境信号通路的蛋白表达减少;淫羊藿总黄酮可改善小肠干细胞的增殖分化功能,上调干细胞微环境Wnt/β-catenin和BMP信号通路相关蛋白表达。

骨髓基质细胞对中脑神经干细胞分化为神经元的诱导作用

目的:观察成年大鼠骨髓基质细胞诱导新生大鼠中脑神经干细胞分化为神经元的机制。方法:采用骨髓基质细胞和神经干细胞共培养方法,通过显微镜观察神经干细胞的分化状态;使用免疫组织化学技术,分析神经元在神经干细胞后代中所占的比例。结果:(1)骨髓基质细胞可诱导神经干细胞分化为高比例神经元;(2)骨髓基质细胞可促进神经元的存活。结论:骨髓基质细胞可提供神经干细胞分化为神经元和促进神经元存活的信号物质。

胚胎干细胞向造血干/祖细胞定向诱导分化的研究进展

胚胎干细胞(embryonic stem cell,ES细胞)是指由胚胎内细胞团(inner cell mass,ICM)细胞经体外抑制培养而筛选得到的细胞,具有无限增殖潜能,在体外可以向造血细胞分化,有可能为造血干细胞移植和血细胞输注开辟新的来源.此外,ES细胞向造血干/祖细胞的定向诱导分化也为阐明哺乳动物造血发育的细胞和分子机制提供了良好的体外模型.对ES细胞向造血干/祖细胞定向分化的研究进展进行了综述.

神经干细胞移植对大鼠视神经损伤后节细胞的保护作用

目的探讨神经干细胞(NSCs)移植对视神经受损SD大鼠视网膜节细胞(RGCs)的保护作用。方法将48只健康成年SD大鼠随机分为N组(NSCs移植组)和C组(对照组),均使用精确校准方法在右眼造成部分视神经损伤,左眼作为正常对照。从胚胎SD大鼠海马分离NSCs,利用细胞培养和体内移植技术,将培养后的NSCs注入N组大鼠右眼玻璃体内,C组大鼠右眼玻璃体内注入同等体积的PBS,处死前3d在双上丘注射3%快蓝逆行标记双眼RGCs。将大鼠分别于注射NSCs或PBS后7d、14d、21d、28d处死,各分为4组,每组6只,分离视网膜置于荧光显微镜下,摄影输入计算机图像分析仪计数RGC,计算RGC标识率。另取6只健康成年SD大鼠作NSCs移植,分别于移植后7d、28d处死,每时间段3只,通过视网膜免疫荧光切片观察NSCs在视网膜的存活、整合情况。结果N组大鼠各时间段RGCs标识率与C组同时间段比较均增高,有显著性差异(P<0.01);N组与C组RGCs标识率均随时间延长而降低,且前后段时间比较有显著性差异,但N组降低速度明显慢于C组。NSCs移植7d后部分移植细胞迁移进入视网膜的内网层和节细胞层,28d后可见移植细胞广泛整合至宿主视网膜内。结论NSCs移植入视神经损伤大鼠视网膜后可提高视网膜神经节细胞的存活率,对受损的节细胞具有一定的保护作用。

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

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分数值有显著改善,差异有显著性意义(P<0.05);(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(P<0.05);(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。