兰州大学机构库 >学院待认领
心肌注射骨髓间充质基质细胞后逃逸细胞对心肌梗死大鼠模型心外脏器的影响
Alternative TitleImpact of escaped bone marrow mesenchymal stromal cells on extra-cardiac organs after intramyocardial implantation in a rat myocardial infarction model
王炜
Thesis Advisor高秉仁
2010-05-21
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword骨髓间充质基质细胞 心肌梗死 肺脏 肝脏 脾脏
Abstract目的: 移植骨髓来源的间充质基质细胞(Mesenchymal stromal cells, MSCs)已成为心肌梗死治疗的新热点。心肌注射已经成为细胞精确输注至目标区心肌的最有效的途径。然而,经心肌注射细胞治疗心肌梗死后会发生细胞逃逸,逃逸的细胞随之会被心外脏器特别是体循环和肺循环的过滤器官所截留。我们设计此试验来探索迁移的骨髓来源的间充质基质细胞的体内分布、最终命运以及其对肺脏、肝脏和脾脏的影响。 方法: MSCs取自雄性Lewis大鼠,冠状动脉结扎法制作雌性Lewis大鼠心肌梗死(Myocardial infarction, MI)模型。心梗3周后,经超声筛选左室射血分数小于60%同时左室缩短分数小于30%的心肌梗死大鼠被随机分两组,细胞移植组(n=22)于心梗边缘区单点注射5-溴脱氧尿嘧啶核苷(BrdU)标记的雄性MSCs3×106个/50 μl;对照组(n=21)注射相同体积的磷酸盐缓冲液(PBS)。10只健康大鼠作为假手术组只接受心肌内注射相同体积的PBS。MSCs移植后4周,超声心动图评价左心功能,采血测定肝功能、血常规及免疫球蛋白M水平,采用酶联免疫吸附法(ELISA)及比色法分别测定肺组织中肿瘤坏死因子-α(Tumour necrosis factor-α,TNF-α),白细胞介素-1(interleukin-1β,IL-1β)含量及髓过氧化物酶(myeloperoxidase,MPO)活性,应用实时定量聚合酶链反应来测定各脏器中的细胞存留(real time-polymerase chain reaction,qRT-PCR),免疫组织化学双染法检测移植细胞及其分化。 结果: 细胞移植后心功能得到改善。qRT-PCR结果显示移植细胞在心脏中存留数量最多,其次依次为脾脏、肝脏和肺脏。肝功能、脾功能及组织匀浆检测均未发现细胞移植造成了明显的副反应。相反,细胞移植组大鼠血清白蛋白水平明显升高,谷丙转氨酶水平明显下降,提示细胞移植在一定程度上改善了心梗大鼠的肝脏功能。移植后逃逸至肝脏、脾脏和肺脏的细胞未分化为成纤维细胞、肌成纤维母细胞或者肺泡上皮细胞。同时,我们观察到逃逸到肝脏的移植细胞表达了功能性肝细胞的标志物。 结论:经心肌注射MSCs后发生的移植细胞迁移未导致肺脏、肝脏和脾脏功能的恶化。然而,其他干细胞源治疗心肌梗死时对心外脏器的生物安全性问题仍然需要进一步探索。
Other AbstractObjectives: Recent years have witnessed a growing interest and enthusiasm in the application of bone-marrow derived stromal cell-based therapies to repair or regenerate damaged myocardium. Intramyocardial injection has become the most effective route of cell infusion precisely to the target area. Cell escape would occur after intramyocardial injection for the treatment of myocardial infarction (MI) and then the migrated cells would be entrapped by the extra-cardiac organs especially by the filter organs of the systemic and pulmonary circulation. We investigated the distribution and fate of injected bone marrow-derived mesenchymal stromal cells (MSCs) and their impact on the lung, liver and spleen. Materials and Methods: MSCs were isolated from male donor Lewis rat. Myocardial infarction was induced by left anterior descending artery ligation in female Lewis rats. Three weeks later, the qualified female rats with left ventricular ejection fraction (LVEF) lower than 60% and left ventricular fractional shortening (LVFS) lower than 30% were randomly divided for 2 groups. In cell transplantation group, 3 millions male MSCs or phosphate buffered solution was injected into infarcted myocardium. 10 healthy rats received intramyocardial PBS injection and were served as sham group. Four weeks after cell transplantation, left ventricular function were evaluated.The blood was obtained for the assessment of liver function, routine blood test and the level of immunoglobulin M. Tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) level in lung were measured with antibody enzyme-linked immunosorbent assays (ELISA) with commercial ELISA kits. The myeloperoxidase (MPO) activity of lung tissue was measured in colorimetric method. The cell retention was evaluated by quantitative real time-polymerase chain reaction (qRT-PCR). Immunohistochemistry study was performed to identify the migrated cells and differentation. Results: Heart function was improved after cell injection. qRT-PCR results showed the number of retained cells in spleen, liver and lung ranked 2nd ,3rd and 4th followed by the heart without any adverse effects after cell transplantation. Conversely, a significant elevation in serum ALB level and a significant decreasein ALT level in cell transplantation group. It shows partly improve the liver function. The implanted MSCs which escaped to the liver, spleen and lung did not differentiate into fibroblast, myofibroblast or alveolar epithelial cell. Th...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222474
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
王炜. 心肌注射骨髓间充质基质细胞后逃逸细胞对心肌梗死大鼠模型心外脏器的影响[D]. 兰州. 兰州大学,2010.
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