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1.
目的探讨内皮祖细胞(EPC)移植治疗糖尿病兔下肢缺血的病理及缺血肌肉中血管内皮生长因子(VEGF)的变化。方法2006年1月至9月在哈尔滨医科大学附属第二医院内分泌科将30只健康日本大耳白兔随机分为3组,即糖尿病磷酸盐缓冲液(PBS)对照组(A组)8只、糖尿病内皮祖细胞移植治疗组(B组)14只、正常血糖内皮祖细胞移植治疗组(C组)8只。骨髓来源的内皮祖细胞经体外扩增培养7d后,通过肌内注射进行细胞移植,用病理改变及肌浆中VEGF的变化评价治疗效果。结果EPC移植后14d,病理示3组毛细血管密度分别为(9.29±1.63)个/视野、(12.60±2.16)个/视野、(12.51±1.56)个/视野;血管数/肌束数分别为0.66±0.05,0.83±0.11,0.90±0.13;肌浆中VEGF质量分数[每克肌肉中VEGF因子的质量(ng)]分别为0.22±0.05,0.30±0.07,0.31±0.08;糖尿病内皮祖细胞移植治疗组与糖尿病PBS对照组比较,毛细血管密度、血管数/肌束数、VEGF值差异均有显著性意义,P<0.05。结论EPC移植能有效治疗糖尿病兔下肢缺血。  相似文献   

2.
目的研究内皮祖细胞(EPC)移植治疗糖尿病兔下肢缺血对血管生成相关基因表达的影响。方法日本大耳白兔16只,四氧嘧啶法建立糖尿病模型后,随机分为磷酸盐缓冲液(PBS)对照组与EPC治疗组,分别用PBS和骨髓来源经体外扩增的EPC移植于下肢缺血区,14天后用基因芯片分析EPC移植对血管生成相关基因表达的影响,同时检测肌浆中血管内皮生长因子(VEGF)浓度。结果EPC治疗组下肢缺血区27个血管生成基因表达上调超过2倍,移植治疗组VEGF的浓度显著高于对照组,差异有统计学意义。结论EPC植入糖尿病兔下肢缺血区,可以上调包括VEGF在内的多种血管生成相关基因,同时肌浆中VEGF水平升高。  相似文献   

3.
目的研究脐血内皮祖细胞(EPC)治疗糖尿病下肢缺血的有效性及机制,为临床治疗糖尿病足病(DF)提供理论依据。方法取产妇足月产脐血分离单核细胞,培养7d,流式细胞仪鉴定细胞,并计数活细胞数。将Wistar雄性大鼠分成3组:(1)糖尿病大鼠射线照射后结扎双后肢股动脉,尾静脉注射血管EPC(DE组)。(2)糖尿病大鼠结扎双后肢股动脉,尾静脉注射缓冲液(DP组)。(3)正常大鼠射线照射后结扎双后肢,尾静脉注射EPC(NC组)。用绿色荧光(GFP)示踪EPC,Ⅷ因子免疫组化检测肌纤维间毛细血管数,RT-PCR检测双后肢肌肉血管内皮生长因子(VEGF)mRNA水平。结果 DE、NC组右后肢腓肠肌溃疡及缺血明显好转,有明显荧光,肌纤维间毛细血管数多,VEGF表达高,DP组好转不明显,无荧光,毛细血管数少,VEGF表达少,其差异有统计学意义(P<0.05)。结论脐血EPC治疗糖尿病大鼠下肢缺血有效。  相似文献   

4.
目的观察不同数量的内皮祖细胞移植对内膜球囊损伤血管修复的影响。方法密度梯度离心法分离兔外周血单个核细胞,以EGM-2培养基培养7天,获得兔外周血内皮祖细胞。分高细胞数量移植组(5×105个细胞)和低细胞数量移植组(2×105个细胞),将相应数量的内皮祖细胞重悬于100μL生理盐水中移植至兔内膜球囊损伤血管局部,对照组仅注入100μL生理盐水。同时用CM-DiI标记内皮祖细胞移植,进行细胞示踪。4周后,处死动物,荧光显微镜下观察CM-DiI标记细胞的分布,并测量各组内膜损伤血管再生内皮覆盖率和新生内膜增生程度。结果荧光显微镜下发现荧光标记的内皮祖细胞分布在血管的中膜层、新生内膜层和内膜表面。内皮祖细胞移植可显著促进内膜损伤血管的再内皮化,以高细胞数量移植组为著,同时内皮祖细胞移植也大大减少了新生内膜的形成。结论内皮祖细胞移植可修复内膜球囊损伤血管,高细胞数量移植有更显著的效果。  相似文献   

5.
目的 观察外周血内皮祖细胞治疗下肢缺血性疾病的疗效,比较内皮祖细胞两种不同移植方式的治疗效果.方法 2008年1月至2010年1月在我院治疗30例患者的34条缺血的下肢,选择两种移植方法,16例18条患肢采用小腿肌肉局部注射,16例17条患肢采用下肢动脉腔内注射.结果 二组患者总的疼痛缓解改善率分别为83.3%(15/18)和93.8%(15/16).对缓解或改善患者的冷、凉感觉,二组患者总的有效率均为100% (16/16,14/14).二组患者总的保肢率分别为88.9%(16/18)和93.75%(15/16).术后两种移植方法的踝肱比值(ABI)增加的例数分别为38.8.4% (8/18)和37.5.2% (6/16).有15例患者16条患肢接受了下肢动脉造影,发现均有不同程度的新生侧枝血管形成.结论 内皮祖细胞移植治疗下肢缺血性疾病是一种简单、安全、有效的方法.两种移植方法疗效没有明显的差异.  相似文献   

6.
目的:探讨移植内皮祖细胞(EPCs)对兔心肌梗死后心功能的影响。方法:通过密度梯度离心法体外分离兔单个核细胞,接种于人纤连蛋白包被的培养板上,并给予含血管内皮生长因子(VEGF)20μg/L的EBM-2完全培养液培养2周。取健康大耳白兔,结扎左冠状动脉前降支,并将制作心肌梗死模型成功的大耳白兔随机分为两组,即对照组和实验组,每组20只。2周后实验组心外膜下直接注射EPCs(1.6×1010/L)悬液,对照组注射无菌PBS液。此外,两组均给予心肌梗死后常规药物治疗。术后4周,分别行超声心动图检查和血浆脑钠肽(BNP)浓度测定及计数移植区毛细血管。结果:实验组左室舒张末期内径(LVEDD)与术前及对照组相比均无统计学差异,而实验组左室射血分数(LVEF)与术前及对照组相比有统计学差异(P0.05)。实验组血浆BNP水平较术前降低,差异有统计学意义(P0.01),且与对照组相比,血浆BNP水平明显降低(P0.05)。移植细胞4周后,实验组梗死边缘区毛细血管密度与对照组有显著性差异(P0.05)。结论:心外膜下直接注射移植EPCs能够改善兔心肌梗死后的心功能,延缓慢性心力衰竭的发生和发展。  相似文献   

7.
一些实验研究表明,内皮祖细胞(EPC)在脑梗死后血管新生中起重要作用。EPC移植有可能为缺血性脑血管病的治疗提供一种全新的治疗策略。文章阐述了新血管形成和EPC与脑缺血的关系,并探讨了EPC移植治疗脑缺血的影响因素和应用前景。  相似文献   

8.
内皮祖细胞是内皮细胞的前体细胞,是成年个体中与血管新生关系最为紧密的干细胞成分,大量动物实验和初步临床研究均显示其在治疗缺血性心脏病、外周缺血性血管疾病及血管损伤后再狭窄等方面具有广阔的临床应用前景。现主要对近年来内皮祖细胞移植策略做一综述。  相似文献   

9.
糖尿病血管并发症严重影响患者的生存质量,甚至威胁生命.近年研究发现,内皮祖细胞(EPCs)的变化可能是糖尿病及其血管并发症发生的重要原因.本文对EPCs减少和功能异常在各种糖尿病血管并发症中的作用及可能的干预治疗的前景作一综述.  相似文献   

10.
内皮祖细胞移植治疗大鼠急性心肌梗死的实验研究   总被引:1,自引:0,他引:1  
目的探讨大鼠内皮祖细胞(EPCs)移植于梗死心肌的增殖分化情况及对心功能的影响。方法分离培养大鼠EPCs,免疫荧光法检测其CD34+、CD133+和Flk-1+的表达。将SD大鼠冠状动脉左前降支结扎制造急性心肌梗死模型后,在梗死心肌处植入DAPI标记的EPCs(实验组)或M 199培养液(对照组)。移植后1周及4周,心脏超声检查心功能,并对梗死区心肌组织进行移植细胞形态学检查及毛细血管密度测定,逆转录聚合酶链反应及酶联免疫吸附测定梗死周边区血管内皮生长因子(VEGF)的表达。结果培养获得EPCs,其表型为CD34+、CD133+和Flk-1+。植入梗死区的EPCs可以分化为血管内皮细胞,实验组梗死心肌处血管密度较对照组明显增高(P<0.01),并且VEGF基因及蛋白表达在移植后1周均较对照组明显增高(P<0.01)。移植后4周,实验组大鼠左心室射血分数及左心室短轴缩短率较对照组明显提高(P<0.01);而移植后1周,两组大鼠超声检测心功能各指标变化不明显。结论同种异体EPCs移植到梗死心肌大鼠缺血心肌能分化为毛细血管内皮细胞,促进梗死后心肌血管新生,改善心功能。  相似文献   

11.
目的比较内皮祖细胞(EPC)和单个核细胞(MNC)移植改善糖尿病大鼠后肢血管病变的疗效。方法将大鼠EPC和外周血MNC标记后分别植入糖尿病大鼠缺血后肢。对照组仅植入PBS溶液。ELISA法检测VEGF的含量,免疫组化法计数毛细血管数目。结果EPC和MNC均能在缺血后肢分化为毛细血管。移植后EPC组和MNC组大鼠缺血肢体局部VEGF含量高于对照组,毛细血管数目也高于对照组。但EPC组和MNC组VEGF含量和毛细血管数目无明显差异。结论移植EPC及含有相当数量EPC的MNC对糖尿病大鼠后肢血管病变的改善作用相似。EPC可能在外周血单个核细胞移植中发挥主要作用。  相似文献   

12.

Background

This study tested the hypothesis that tissue plasminogen activator (tPA) is crucial for regulating endothelial progenitor cell (EPC) mobilization from bone marrow to circulation in murine critical limb ischemia (CLI) by ligating the left femoral artery.

Methods

Wild-type (C57BL/6) (n = 40) mice were equally divided into group 1A (sham control), group 2A (CLI), group 3A [control-tPA (4.0 mg/kg)] and group 4A [CLI-tPA (intravenously at 3 h after CLI)]. Similarly, tPA knock-out (tPA−/−) mice (n = 40) were equally divided into group 1B (sham control), group 2B (CLI), group 3B [control-tPA (4.0 mg/kg)], and group 4B (CLI-tPA).

Results

The circulating levels of EPCs (C-kit/CD31 +, Sca-1/KDR +, CXCR4/CD34 +) were lower in groups 1B and 2B than in groups 1A and 2A, respectively (all p < 0.01), and were reversed after tPA treatment (3B vs. 3A or 4B vs. 4A, p > 0.05) at 6 h and 18 h post-CLI. Levels of these biomarkers decreased again 14 days after CLI in tPA−/− mice compared to those in wild-type between the respective groups (all p < 0.01). Laser Doppler flowmetry showed a higher ratio of ischemic-to-normal blood flow in 2A than in 2B and in 4A than in 4B by day 14 after CLI (all p < 0.05). Angiogenesis at protein (CXCR4, SDF-1α, VEGF) and cellular (CXCR4 +, SDF-1α +, and CD31 + cells) levels was highest in animals with CLI-tPA, significantly higher in mice with CLI only than in sham controls for both wild-type and tPA−/− mice (p < 0.01).

Conclusion

tPA played an essential role in augmenting circulating EPCs, angiogenesis, and blood flow in the ischemic limb in a murine model.  相似文献   

13.
血管内皮祖细胞在治疗动脉粥样硬化中的作用   总被引:1,自引:0,他引:1  
<正>1997年,Asahara等[1]首次发现人体外周血中存在能分化为血管内皮细胞的前体细胞,将其命名为血管内皮祖细胞(endothelial progenitor cells,EPCs)。随后,人们对EPCs进行了大量的研究。内皮祖细胞不仅参与胚胎期的血管发生,而且在成体的血管生长发育中发挥重要作用,对于创伤修  相似文献   

14.
15.
万钧  王辰  王军 《国际呼吸杂志》2009,29(22):1393-1396
内皮祖细胞是成熟血管内皮细胞的前体细胞,属于干细胞群体,其在血管再生与修复方面的应用越来越受到关注.肺动脉高压作为一种致死性很高的肺血管疾病,其发病与血管内皮损伤密切相关.目前的研究证实,内皮祖细胞在肺动脉高压血管内皮修复中具有重要作用,内皮祖细胞移植有可能成为临床治疗肺动脉高压的一种有效手段.  相似文献   

16.
Circulating endothelial progenitor cells (EPC) are incorporated into newly formed capillaries, enhance neovascularization after hind limb ischemia and improve cardiac function after ischemic injury. Incorporated progenitor cells may also promote neovascularization and cardiac regeneration by releasing factors, which act in a paracrine manner to support local angiogenesis and mobilize tissue residing progenitor cells. Therefore, we analyzed the expression profile of cytokines in human peripheral blood-derived EPC as opposed to human umbilical vein endothelial cells (HUVEC), human microvascular endothelial cells (HMVEC), and CD14(+) monocytes by microarray technology. A gene tree analysis revealed a distinct expression pattern of angiogenic growth factors in EPC, mature endothelial cells, and CD14(+) monocytes. VEGF-A, VEGF-B, SDF-1, and IGF-1 mRNA levels were higher in EPC as compared to HUVEC or HMVEC. The enhanced mRNA expression was paralleled by a significant release of VEGF, SDF-1, and IGF-1 protein into the cell culture supernatant of EPC. Moreover, immunohistological analysis of ischemic limbs from nude rats revealed that VEGF is also released from recruited human EPC in vivo. As a functional consequence, conditioned medium of EPC induced a strong migratory response of mature endothelial cells, which was significantly inhibited by VEGF and SDF-1 neutralizing antibodies. Finally, conditioned medium of EPC significantly stimulated the migration of cardiac resident c-kit(+) progenitor cells in vitro. Taken together, EPC exhibit a high expression of angiogenic growth factors, which enhanced migration of mature endothelial cells and tissue resident cardiac progenitor cells. In addition to the physical contribution of EPC to newly formed vessels, the enhanced expression of cytokines may be a supportive mechanism to improve blood vessel formation and cardiac regeneration after cell therapy.  相似文献   

17.
兔血管内皮祖细胞的分离及诱导分化   总被引:1,自引:2,他引:1  
目的:探讨自兔骨髓中分离及诱导血管内皮祖细胞(EPCs)的方法。方法:抽取兔骨髓,Ficoll离心液梯度离心,分离出骨髓单个核细胞,用含有血管内皮生长因子(VEGF)、碱性纤维细胞生长因子(bFGF)、胰岛素样生长因子-1(IGF-1)的M199培养液诱导培养EPCs,透射电镜及免疫荧光法对所培养细胞进行鉴定。结果:细胞诱导培养48~72h后即可见有细胞贴壁,逐渐变成梭形。培养至8d左右,可观察到数条由梭形贴壁细胞直线生长连接而成的条索,7~10d出现多个细胞团,贴壁的梭形细胞开始从细胞团的边缘出芽长出,培养至14d左右,细胞逐渐融合,连接成大片条索状结构。透射电镜及免疫荧光检查证实为血管内皮祖细胞。结论:梯度离心法自兔骨髓获得单个核细胞,在体外经诱导分化可获得血管内皮祖细胞。  相似文献   

18.
Augmenting neovascularization with the use of endothelial progenitor cells (EPCs) is a therapeutic option to rescue critical limb ischemia (CLI). However, the outcomes have been not so satisfactory. The detectable number of injected EPCs at the ischemic site is rather small. If EPCs accumulate more and stay longer there, neovascularization will be augmented. In this study, we tested whether external magnetic forces (EMFs) accumulated magnetized EPCs (Mag-EPCs) at the ischemic site and thereby augmented neovascularization. We cultured peripheral blood-derived mononuclear cells to obtain EPCs and generated Mag-EPCs by a magnetofection method with nanoparticles. Prussian-blue staining revealed magnetic nanoparticles incorporated into the cytoplasms and nuclei of Mag-EPCs. The survival rate of Mag-EPCs at day 9 of culture was 98.7%, indicating no cell toxicity of magnetic nanoparticles. EMFs augmented adhesion capacity of Mag-EPCs not only in the static but also in the flow condition in vitro, compared to without EMFs. The migration capacity of Mag-EPCs with EMFs was 160% more than EPCs or Mag-EPCs without them. After an intravenous injection of Mag-EPCs into the rat with hind-limb ischemia, the recovery of blood flow and capillary density in the ischemic limb were significantly more (p < 0.01) with EMFs than without them. EMFs augmented neovascularization capacity of Mag-EPCs compared to EPCs alone. This method could be a new therapeutic strategy for patients with CLI.  相似文献   

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