Irradiated cultured apoptotic peripheral blood mononuclear cells regenerate infarcted myocardium
Irradiated cultured apoptotic peripheral blood mononuclear cells regenerate infarcted myocardium
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DOI：
10.1111/j.1365-2362.2009.02111.x
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发表时间：
2009-06-01
影响因子：
5.5
通讯作者：
Podesser, B. K.
中科院分区：
文献类型：
--
作者：
Ankersmit, H. J.;Hoetzenecker, K.;Podesser, B. K.
急性心肌梗死（AMI）后发生心肌重塑，常导致充血性心力衰竭。为研究c-kit+内皮祖细胞（EPC）归巢对梗死心肌再生的作用，采用体外培养的方法，观察了活化的外周血单个核细胞（PBMC）与辐射凋亡PBMC（IA-PBMC）共培养后的免疫功能。24小时后获得活PBMC、IA-PBMC及其培养上清液（SN）。采用逆转录聚合酶链反应（RT-PCR）和酶联免疫吸附法（ELISA）检测外周血单个核细胞（PBMC）、黑色素（SN）和SN暴露的成纤维细胞中白细胞介素8（IL-8）、血管内皮生长因子（VEGF）和基质金属蛋白酶9（MMP 9）的含量。将活的和IA-PBMC的细胞悬液输注到实验性大鼠AMI模型中。在梗死后72小时内进行免疫组织化学分析以检测炎性和促血管生成细胞。通过超声心动图和Elastica货车Gieson染色定量功能数据和梗塞大小的测定。IA-PBMC在体外减弱免疫反应性并导致促血管生成IL-8和MMP 9的分泌。暴露于活的和IA-PBMC衍生的SN的成纤维细胞引起IL-8和MMP 9的RNA增加。与对照组（仅培养基，活PBMC）相比，用IA-PBMC细胞悬液输注的AMI大鼠在72小时内证明内皮祖细胞归巢增强。超声心动图显示梗死面积显着减少和改善后AMI重构的衰减损失的射血分数证明。这些数据表明，IA-PBMC细胞悬液在实验性AMI规避炎症，引起优先归巢的再生EPC和替换梗死心肌。
Acute myocardial infarction (AMI) is followed by post AMI cardiac remodelling, often leading to congestive heart failure. Homing of c-kit+ endothelial progenitor cells (EPC) has been thought to be the optimal source for regenerating infarcted myocardium.Immune function of viable peripheral blood mononuclear cells (PBMC) was evaluated after co-culture with irradiated apoptotic PBMC (IA-PBMC) in vitro. Viable PBMC, IA-PBMC and culture supernatants (SN) thereof were obtained after 24 h. Reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay were utilized to quantify interleukin-8 (IL-8), vascular endothelial growth factor, matrix metalloproteinase-9 (MMP9) in PBMC, SN and SN exposed fibroblasts. Cell suspensions of viable- and IA-PBMC were infused in an experimental rat AMI model. Immunohistological analysis was performed to detect inflammatory and pro-angiogenic cells within 72 h post-infarction. Functional data and determination of infarction size were quantified by echocardiography and Elastica van Gieson staining.The IA-PBMC attenuated immune reactivity and resulted in secretion of pro-angiogenic IL-8 and MMP9 in vitro. Fibroblasts exposed to viable and IA-PBMC derived SN caused RNA increment of IL-8 and MMP9. AMI rats that were infused with IA-PBMC cell suspension evidenced enhanced homing of endothelial progenitor cells within 72 h as compared to control (medium alone, viable-PBMC). Echocardiography showed a significant reduction in infarction size and improvement in post AMI remodelling as evidenced by an attenuated loss of ejection fraction.These data indicate that infusion of IA-PBMC cell suspension in experimental AMI circumvented inflammation, caused preferential homing of regenerative EPC and replaced infarcted myocardium.