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1.
目的比较脂肪源间充质干细胞(AMSCs)和骨髓来源间充质干细胞(BMSCs)的免疫调节能力的差异。方法用流式细胞仪分析AMSCs和BMSCs的表型。通过MSCs和淋巴细胞共培养体系,检测MSCs对T细胞增殖、周期、活化、凋亡以及Th细胞分化的影响。结果表型分析结果显示AMSCs和BMSCs的表型基本一致。AMSCs和BMSCs都能抑制T细胞的增殖和早期活化,并在调节辅助性T细胞亚群的分化上作用类似。但在MSC对活化后T细胞的凋亡影响方面,BMSCs能够抑制活化T细胞的凋亡,而AMSC没有这种作用。结论 AMSCs和BMSCs对T淋巴细胞的影响基本类似,其对活化T细胞凋亡影响的差异还有待进一步的机制研究。 相似文献
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Serbay Ozkan Basak Isildar Mahmut Oncul Zafer Baslar Semih Kaleli 《Ultrastructural pathology》2018,42(3):199-210
Mesenchymal stem cells (MSCs) are considered as an important tool for regenerative medicine and experimental treatments. Unveiling the ultrastructural changes during the differentiation of MSCs might help us to understand the nature of the process and to develop novel therapeutic approaches. For this purpose, human umbilical cord (hUC) was chosen as MSC source. In the first place, MSCs were isolated from sub-amniotic, intervascular and perivascular areas of hUC by enzymatic and tissue explant method to determine the most favorable region of hUC and technique for further processing. Therefore, microscopic and growth kinetics analyses showed that there was no clear difference in the morphologies and proliferation rates among the hUC-MSC groups. Flow cytometric analysis showed that CD44 and CD90 MSC markers were highly expressed, while CD34 and CD45 hematopoietic stem cells markers were expressed at low degree. Because our preliminary results showed that there was no conspicuous superiority among the hUC-MSCs groups, whole UC was utilized as a source, and tissue explant method was applied to isolate MSCs for further differentiation analysis. At the 1st and 3rd week of osteogenic and adipogenic differentiation, ultrastructural analysis showed an increase in the number of secondary lysosomes in comparison with the undifferentiated status. Increase in the mitochondrial content was also detected at the 1st week of adipogenic differentiation. Consequently, ultrastructural changes including increase in the number of mitochondria and secondary lysosomes during the adipogenic and osteogenic differentiation could be attributed to the switch in energy metabolism of the MSCs and increment in the lysosomal activity respectively. 相似文献
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目的 探讨骨髓间充质干细胞离体培养并与去抗原牛松质骨复合修复节段性骨缺损的效果。方法 Wistar大鼠骨髓间充质干细胞(MSCs)与去抗原牛松质骨(BCB)制成MSCs/BCB复合物;采用同种异体大鼠桡骨干5mm节段性骨缺损动物模型,通过X线放射学、组织学、生物力学检测对比研究骨缺损修复情况。结果 术后2、4、8、14、18周实验组与实验对照组经放射学检查评价新骨生成有显著性差异。术后组织学检查新骨生成速度、生成量均有显著性差异。18周经生物力学实验检测,实验组与实验对照组新骨生成有显著性差异,实验组标本与正常基本一致。空白对照组各时间点均无新骨形成,最后缺损由纤维组织充填。结论 大鼠骨髓间充质干细胞(MSCs)是骨组织工程中适宜的种子细胞。去抗原牛松质骨是可以选择的细胞载体。MSCs/BCB复合物植入修复骨缺损效果明显优于单纯BCB植入。 相似文献
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目的探讨骨髓间充质干细胞离体培养并与去抗原牛松质骨复合修复节段性骨缺损的效果.方法 Wistar大鼠骨髓间充质干细胞(MSCs)与去抗原牛松质骨(BCB)制成MSCs/ BCB复合物;采用同种异体大鼠桡骨干5mm节段性骨缺损动物模型,通过X线放射学、组织学、生物力学检测对比研究骨缺损修复情况.结果术后2、4、8、14、18周实验组与实验对照组经放射学检查评价新骨生成有显著性差异.术后组织学检查新骨生成速度、生成量均有显著性差异.18周经生物力学实验检测,实验组与实验对照组新骨生成有显著性差异, 实验组标本与正常基本一致.空白对照组各时间点均无新骨形成,最后缺损由纤维组织充填.结论大鼠骨髓间充质干细胞(MSCs)是骨组织工程中适宜的种子细胞.去抗原牛松质骨是可以选择的细胞载体.MSCs/BCB复合物植入修复骨缺损效果明显优于单纯BCB植入. 相似文献
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Liang-Xin Tay Chin-Keong Lim Azura Mansor Tunku Kamarul 《International journal of medical sciences》2014,11(1):24-33
Objective: This preliminary study aims to determine the differentially expressed proteins from chondrogenic differentiated multipotent stromal cells (cMSCs) in comparison to undifferentiated multipotent stromal cells (MSCs) and adult chondrocytes (ACs).Methods: ACs and bone marrow-derived MSCs were harvested from New Zealand White rabbits (n = 3). ACs and cMSCs were embedded in alginate and were cultured using a defined chondrogenic medium containing transforming growth factor-beta 3 (TGF-β3). Chondrogenic expression was determined using type-II collagen, Safranin-O staining and glycosaminoglycan analyses. Two-dimensional gel electrophoresis (2-DE) was used to isolate proteins from MSCs, cMSCs and ACs before being identified using liquid chromatography-mass spectrometry (LC-MS). The differentially expressed proteins were then analyzed using image analysis software.Results: Both cMSCs and ACs were positively stained with type-II collagen and safranin-O. The expression of glycosaminoglycan in cMSCs was comparable to AC at which the highest level was observed at day-21 (p>0.05). Six protein spots were found to be most differentially expressed between MSCs, cMSCs and ACs. The protein spots cofilin-1 (CFL1) and glycealdehyde-3-phosphate dehydrogenase (GAPD) from cMSCs had expression levels similar to that of ACs whereas the others (ie. MYL6B, ALDOA, TAGLN2, EF1-alpha), did not match the expression level of ACs.Conclusion: Despite having similar phenotypic expressions to ACs, cMSCs expressed proteins which were not typically expected. This may explain the reason for the unexplained lack of improvement in cartilage repair outcomes reported in previous studies. 相似文献
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目的 探讨不同炎性因子刺激下细胞因子信号转导抑制分子-1(suppressor of cytokine signaling1,SOCS1)在人骨髓间充质干细胞(mesenchymal stem cells,MSCs)中的表达情况,并研究稳定表达SOCS1的MSCs细胞株的建立方法.方法 用SOCS1相关炎性因子刺激剂处理24小时后裂解MSCs,Westen Blot检测各组细胞SOCS1的表达.通过Gateway技术,构建出表达SOCS1基因的载体质粒pFinal/PGK-puro-EF1 α-SOCS1-IRES-EGFP,将其与包装质粒共转染293FT细胞产生慢病毒,通过多次感染将载体导入人骨髓间充质干细胞,流式细胞术筛选出稳定表达SOCS1的人骨髓间充质干细胞,并用Westen Blotting、流式细胞术、成骨成脂肪诱导分化来鉴定.结果 SOCS1蛋白在IFN-γ、Pam3CSK4、Poly(I∶C)、LPS和ODN 2006刺激的MSCs中表达升高(P<0.01).表达载体包装出的病毒感染后的MSCs中SOCS1蛋白水平提高,流式细胞检测表达(>95%)CD29、CD44、CD73、CD90、CD105、CD166,不表达(<2%)CD34和CD45,在相应诱导剂诱导下可分化为成骨细胞和脂肪细胞.结论 多种炎性因子能诱导MSCs内SOCS1表达升高,说明SOCS1可能参与了MSCs的免疫调节功能;成功建立了稳定表达SOCS1的MSCs细胞株. 相似文献
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在皮肤组织工程种子细胞的研究中,表皮干细胞在皮肤再生过程中起了至关重要的作用。但是,对表皮干细胞或前体细胞的分离和扩增水平有限,限制了其临床应用。间充质干细胞(mesenchymal stem cell,MSCs)可诱导分化为皮肤表皮细胞和血管内皮细胞,并具有独特的生物学特性,如来源广泛、易分离培养扩增及免疫源性低等,使其可能成为皮肤组织工程中的种子细胞,为皮肤缺损临床治疗另辟新径。就MSCs治疗皮肤缺损的临床应用作一概述。 相似文献
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《Biomaterials》2015
Implantation of scaffolds may elicit a host foreign body response triggered by monocyte/macrophage lineage cells. Growing evidence suggests that topographical cues of scaffolds play an important role in MSC functionality. In this work, we examined whether surface topographical features can regulate paracrine interactions that MSCs establish with macrophages. Three-dimensional (3D) topography sensing drives MSCs into a spatial arrangement that stimulates the production of the anti-inflammatory proteins PGE2 and TSG-6. Compared to two-dimensional (2D) settings, 3D arrangement of MSCs co-cultured with macrophages leads to an important decrease in the secretion of soluble factors related with inflammation and chemotaxis including IL-6 and MCP-1. Attenuation of MCP-1 secretion in 3D co-cultures correlates with a decrease in the accumulation of its mRNA levels in MSCs and macrophages. Using neutralizing antibodies, we identified that the interplay between PGE2, IL-6, TSG-6 and MCP-1 in the co-cultures is strongly influenced by the micro-architecture that supports MSCs. Local inflammatory milieu provided by 3D-arranged MSCs in co-cultures induces a decrease in monocyte migration as compared to monolayer cells. This effect is partially mediated by reduced levels of IL-6 and MCP-1, proteins that up-regulate each other's secretion. Our findings highlight the importance of topographical cues in the soluble factor-guided communication between MSCs and macrophages. 相似文献
9.
目的 检测比较间充质干细胞(MSCs)移植治疗对烧伤大鼠炎症相关指标白细胞(WBC)、C反应蛋白(CRP)、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)水平变化的影响,探讨MSCs移植治疗烧伤的抑炎机制、效果及临床意义.方法 取正常新生儿脐带,分离脐带华通氏胶,培养脐带MSCs.另取体质量为(200±5)g SD雄性大鼠制备烧伤模型并随机分为对照组和移植组,移植组在烧伤24 h后进行皮下注射MSCs移植(2×106个/只).分别于烫伤后0、1、2、3、5、7d采集血样,检测其WBC、CRP及炎症因子IFN-γ、TNF-α、IL-6、IL-10含量,并比较7、14、21、28 d时2组间创面愈合率及创面愈合时间.结果 对照组WBC烧伤后1、2d显著升高,3d后逐渐降低,移植组仅出现轻微升高;CRP水平变化比较,对照组伤后2、3d出现较大幅度上升,移植组变化甚微;血清中的炎症因子TNF-α、IL-6和IL-10比较,对照组和移植组于伤后3、5、7d均有增高,其中对照组增高更为明显;IFN-γ比较2组间未见明显差别.MSCs移植组14、21、28 d创面愈合率及创面愈合时间均明显优于对照组.结论 MSCs移植治疗烧伤大鼠,能明显减少烧伤大鼠血清中WBC、CRP、TNF-α、IL-6、IL-10的含量,减轻了由细胞炎症因子介导的炎症反应,但MSCs移植并未影响IFN-γ的产生.MSCs移植可通过降低烧伤后炎症因子产生而抑制其继发性炎症反应,从而促进创面愈合及烫伤修复. 相似文献
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The response of human bone marrow derived human mesenchymal stem cells (hMSCs) encapsulated in silk ionomer hydrogels was studied. Silk aqueous solutions with silk-poly-l-lysine or silk-poly-l-glutamate were formed into hydrogels via ultrasonication in situ with different net charges. hMSCs were encapsulated within the hydrogels and the impact of matrix charge was assessed over weeks in osteogenic, adipogenic and maintenance growth media. These modified silk charged polymers supported cell viability and proliferative potential, and the hMSCs were able to differentiate toward osteogenic or adipogenic lineages in the corresponding differentiation media. The silk/silk-poly-l-lysine hydrogels exhibited a positive effect on selective osteogenesis of hMSCs, inducing differentiation toward an osteogenic lineage even in the absence of osteogenic supplements, while also inhibiting adipogenesis. In contrast, silk/silk fibroin-poly-l-glutamate hydrogels supported both osteogenic and adipogenic differentiation of hMSCs when cultured under induction conditions. The results demonstrate the potential utility of silk-based ionomers in gel formats for hMSCs encapsulation and for directing hMSCs long term functional differentiation toward specific lineages. 相似文献
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12.
目的探讨骨髓源间充质干细胞(mesenchymal stem cells,MSCs)对移植物抗宿主病(graft versus host disease,GVHD)模型大鼠的免疫调节作用。方法从SD大鼠骨髓中分离培养MSCs。通过给放射线照射过的Wistar大鼠进行干细胞和单个核细胞(mononuclear cell,MNC)共同移植,建立大鼠GVHD模型,对其进行异基因骨髓移植的同时,共输注不同数量的供者源性的MSCs,观察受鼠的一般表现、生存期及病理损伤的变化;利用细胞免疫化学检测各实验组大鼠外周血MNC中Foxp3+细胞比例的变化,应用ELISA方法检测实验各组大鼠外周血血清中细胞因子IFN-γ、IL-4的变化,诱导各组受鼠脾脏MNC为CTL,并用MTT法检测各组受鼠CTL的杀伤活性。结果异基因骨髓移植时,共输注供鼠MSCs能够明显降低GVHD的发生率,并减轻其病理损伤,延长生存时间;共输注MSCs受鼠血清中IFN-γ水平明显减低而IL-4水平明显增加(P<0.05),与GVHD组大鼠相比,共输注MSCs的各组大鼠外周血中Foxp3+细胞比例明显增加(P<0.05),而CTL杀伤活性明显减弱。结论 MSCs能够通过免疫调节作用抑制GVHD的发生,降低其严重程度,并延长受鼠生存时间。 相似文献
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目的研究大鼠骨髓间充质干细胞(MSCs)体外向心肌样细胞分化过程中L-型钙通道(α1C)和瞬间外向钾通道(kv4.3)基因的表达。方法无菌条件下冲洗Wistar大鼠双侧股骨和胫骨的骨髓腔获得MSCs,体外培养传代纯化,5-氮杂胞苷(5-aza)诱导24 h,显微镜下观察诱导前后形态变化,RT-PCR鉴定α1C和kv4.3基因的表达。结果kv4.3基因诱导前有弱的表达,诱导后1、4、7和14 d表达明显逐渐增强(P<0.05);α1C基因诱导前有较强的表达,诱导后1、4、7和14 d表达却明显逐渐减弱(P<0.05)。结论α1C和kv4.3基因在维持MSCs的诱导分化过程中发挥重要作用。 相似文献
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目的探索葛根素体外诱导大鼠骨髓间充质细胞(Bone Marrow Stromal Cells MSCs)分化为神经元和胶质细胞的可行性,为中药在细胞定向分化中的应用提供理论和实验依据.方法用贴壁法分离纯化SD大鼠骨髓间充质细胞.培养传至第5代,诱导组以2.5mg/ml的葛根素预诱导24h后,用17.5mg/ml的葛根素无血清培养基诱导,未诱导组加入等量培养基,24 h后相差显微镜观察形态变化,免疫细胞化学染色鉴定诱导后的细胞神经元特异性稀醇化酶(neuronspecific enolase NSE),神经胶质纤维酸性蛋白(glial fibrillary acidic protein GFAP)蛋白的表达情况,MTT检测不同浓度葛根素诱导后细胞的活力,流式细胞仪及RT-PCR检测诱导前后细胞中NSE、GFAP的表达情况.结果诱导18h后BMSCs胞体收缩,有突起伸出,24 h后突起增多呈网状.免疫细胞化学染色,NSE阳性表达率为(53.3±4.3)%,GFAP阳性表达为(64.5±5.2)%,流式细胞仪检测诱导24 h后的细胞NSE及GFAP表达量均较未诱导组升高,RT-PCR检测诱导后细胞表达NSE、GFAP,未诱导的细胞则不表达.结论葛根素可诱导大鼠骨髓间充质细胞在体外分化为神经元和神经胶质细胞. 相似文献
15.
Li W Ma N Ong LL Nesselmann C Klopsch C Ladilov Y Furlani D Piechaczek C Moebius JM Lützow K Lendlein A Stamm C Li RK Steinhoff G 《Stem cells (Dayton, Ohio)》2007,25(8):2118-2127
Engraftment of mesenchymal stem cells (MSCs) derived from adult bone marrow has been proposed as a potential therapeutic approach for postinfarction left ventricular dysfunction. However, limited cell viability after transplantation into the myocardium has restricted its regenerative capacity. In this study, we genetically modified MSCs with an antiapoptotic Bcl-2 gene and evaluated cell survival, engraftment, revascularization, and functional improvement in a rat left anterior descending ligation model via intracardiac injection. Rat MSCs were manipulated to overexpress the Bcl-2 gene. In vitro, the antiapoptotic and paracrine effects were assessed under hypoxic conditions. In vivo, the Bcl-2 gene-modified MSCs (Bcl-2-MSCs) were injected after myocardial infarction. The surviving cells were tracked after transplantation. Capillary density was quantified after 3 weeks. The left ventricular function was evaluated by pressure-volume loops. The Bcl-2 gene protected MSCs against apoptosis. In vitro, Bcl-2 overexpression reduced MSC apoptosis by 32% and enhanced vascular endothelial growth factor secretion by more than 60% under hypoxic conditions. Transplantation with Bcl-2-MSCs increased 2.2-fold, 1.9-fold, and 1.2-fold of the cellular survival at 4 days, 3 weeks, and 6 weeks, respectively, compared with the vector-MSC group. Capillary density in the infarct border zone was 15% higher in Bcl-2-MSC transplanted animals than in vector-MSC treated animals. Furthermore, Bcl-2-MSC transplanted animals had 17% smaller infarct size than vector-MSC treated animals and exhibited functional recovery remarkably. Our current findings support the premise that transplantation of antiapoptotic gene-modified MSCs may have values for mediating substantial functional recovery after acute myocardial infarction. 相似文献
16.
目的:观察胎鼠间充质干细胞(MSCs)缓解体力疲劳的作用,探讨其改善肌无力患者或者中老年体弱者生活质量的意义。方法:取孕13.5 d的BALB/c胎鼠,分离纯化得到胎鼠MSCs。6月龄BALB/c雌性小鼠随机分为安静对照组、单纯运动组和MSCs+运动组。MSCs+运动组尾静脉输注第5代MSCs, 其余2组输注同体积生理盐水。24 h后进行负重游泳试验,测定血尿素氮(BUN)、血乳酸、肝糖原和肌糖原,以及肝、肌肉、脑组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,心脏超声检查评价心功能。结果:(1)原代细胞呈梭形、多角形,散在、集落式分布;第5代细胞均匀平行排列或者漩涡状排列,表达CD44和CD105,不表达CD34和CD45,能向成骨细胞或脂肪细胞分化,具备MSCs基本特点。(2)MSCs缓解体力疲劳作用:MSCs+运动组与单纯运动组比较,负重游泳时间明显延长,BUN和血乳酸水平降低,肝糖原和肌糖原储备增加,肝脏和肌肉组织SOD活性升高,MDA水平降低,以上差异均显著(P<0.05)。(3)心功能增强:MSCs+运动组与单纯运动组比较,每搏输出量、心输出量和左室舒张期容积均增加。结论:胎鼠间充质干细胞移植具有缓解小鼠体力疲劳作用。 相似文献
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目的: 观察血管生成素-1(Ang-1)和骨髓间质干细胞(MSCs)对脑梗死的联合治疗作用。方法: 构建带有大鼠Ang-1基因的慢病毒载体,感染大鼠骨髓间质干细胞(rMSCs)获得可表达具有生物活性Ang-1蛋白的rMSCs(Ang-rMSCs)。制备大鼠大脑中动脉栓塞模型,随机分为对照组、Ang-rMSCs组及空载体-rMSCs(pNL-rMSCs)组。脑栓塞术后24 h进行细胞移植。各组大鼠分别于术后24 h、移植后1周、1月及3月用mNSS量表评价神经功能状况,移植后1周应用伊文思蓝(EB)检测血脑屏障通透性,应用荧光激发及荧光免疫组化观察移植细胞在脑内的存活、分化情况,并经组织学观察有无成瘤迹象。结果: 与对照组相比,移植后1周、1月,pNL-rMSCs及Ang-rMSCs组神经功能改善明显(P<0.01),Ang-rMSCs组明显优于pNL-rMSCs组(P<0.001)。移植后3月3组间的mNSS评分差异无显著(P>0.05)。EB检测示血脑屏障通透性在pNL-rMSCs及Ang-rMSCs组低于对照组(P<0.01),Ang-rMSCs组低于pNL-rMSCs组(P<0.01)。移植后1周、1月及3月,Ang-rMSCs和pNL-rMSCs组在荧光激发下均见移植细胞在梗死灶及其周边区聚集并存活。荧光免疫组化检测示Ang- rMSCs组中移植细胞大量表达Ang-1,两移植组中均有部分细胞可同时表达神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF)或星形胶质细胞标志物神经胶质纤维酸性蛋白(GFAP)。未观察到移植细胞异常增殖等成瘤迹象。结论: Ang-1基因修饰的rMSCs移植后可迁移至脑梗死灶周围,分化表达神经细胞标志物并较长期存活。MSCs移植与其分泌的Ang-1蛋白可协同促进脑梗死后神经功能恢复。 相似文献
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MicroRNA regulation associated chondrogenesis of mouse MSCs grown on polyhydroxyalkanoates 总被引:1,自引:0,他引:1
Microbial polyhydroxyalkanoates (PHA) including poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) were found to induce chondrogenesis of mesenchymal stem cells (MSCs) and preserve chondrocytic phenotype as well as support chondrocytes-specific extracellular matrix (ECM) secretion. In this study, mouse MSCs cultured on the PHBHHx films for 24?h showed up-regulated expression of chondrogenic marker genes including aggrecan, col2, sox9, col10 and pthrp. To further illustrate this phenomonon, chondrogenesis-related microRNA expression profiling was examined by quantitative real-time PCR (RT-PCR) based on results of microRNA array obtained from comparison between mouse MSCs and mature mouse chondrocytes. Among 44 microRNAs related to chondrogenesis on microrray studies, considering only broadly-conserved microRNAs, seven differentially-expressed microRNAs were selected to study their target genes related to chondrogenesis. Two microRNAs out of the seven, namely, miR-29a and miR-29b, were revealed to directly target 3' UTR of col2a1 encoding type II collagen by dual-luciferase assay, and their activity was under the regulation of Sox9, the SRY-related high mobility group-box gene 9. For the first time microRNAs were shown to regulate the stem cell differentiation processes mediated by cell-material interactions. 相似文献
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目的 探讨骨髓间充质干细胞 (MSCs) 介导的血红素氧合酶-1 (HO-1) 对心肌梗死后心
脏血管再生及左心室功能的影响。方法 取大鼠骨髓,体外分离扩增培养 MSCs,HO-1 腺病毒转染。结扎左前降支 1 h 后,分别将 HO-1-MSCs、MSCs 多点注射到大鼠心肌梗死区周边,对照组注射等量 PBS。 结果 MSCs 介导的HO-1能在体外及体内获得稳定表达;HO-1-MSCs组促血管生长因子VEGF、FGF2的表达及毛细血管密度明显高于 MSCs 组和对照组 (P < 0.01);但促血管再生的作用可被HO抑制剂阻断。HO-1-MSCs组心肌细胞凋亡及纤维化明显低于MSCs 组和对照组 (P < 0.01);HO-1-MSCs组左室收缩功能各项指标明显优于其他两组(P < 0.01)。HO-1-MSCs组心室壁变厚,心室腔明显缩小。结论 MSCs介导的血红素氧合酶-1能促进心肌梗死后心脏血管再生,改善左心室功能。 相似文献