自体骨髓单个核细胞植入对心肌梗死及其边缘区血管生成相关基因表达的影响

Upregulating the expression of angiogenesis-related genes by transplanting autologous mononuclear bone marrow cells into myocardial infarction scar and the periphery

目的 探讨骨髓单个核细胞植入心肌梗死及其边缘区对血管生成相关基因表达的影响。方法 以冷冻损伤的方法建立新西兰种家兔心肌梗死模型, 4周后,将提取分离的骨髓单个核细胞,植入心肌梗死及其边缘区。以基因芯片研究骨髓单个核细胞植入如何调节心肌梗死及其边缘区血管生成相关基因的表达。并以Westernblot进一步在蛋白水平上验证基因芯片分析结果。结果 骨髓单个核细胞植入心肌梗死及其边缘区后,在心肌梗死区促进 15个血管生成相关基因表达上调超过 3倍,而在边缘区则多达 40个。骨髓单个核细胞植入心肌梗死及其边缘区使血管内皮生长因子、纤维细胞生长因子及血管生长素 Ⅰ等蛋白表达增加,于心肌梗死边缘区作用更加显著。结论 骨髓单个核细胞植入心肌梗死及其边缘区可以上调多种重要血管生成相关基因的表达。

Objective To detect the regulation of angiogenic genes involved in the processes of collateral development Methods Myocardial infarction (MI) scar was induced by cryoinjury in New Zealand rabbits Four weeks after MI, 24 hours before cell transplantation, bone marrow was aspirated from the right thigh bone and mononuclear bone marrow cells (BMCs) were isolated by Ficoll density gradient centrifugation Then the mononuclear BMCs ( n =8) or IMDM culture medium ( n =8) were transplanted into infarction scar and the periphery Four weeks after mononuclear BMCs transplantation, DNA microarray analysis was performed to detect the regulation of angiogenesis related genes in infarction scar and the periphery And the differences of angiogenic genes expression were compared among several important growth factors by Western blot Results DNA microarray analysis showed the detail regulation of genes involved in the angiogenic processes There were 15 genes upregulated over 3 times in the infarction scar In addition, we also found more genes are involved in the process of angiogenesis in its periphery than in the infarction scar (40 genes vs 15 genes) Western bolt analysis further demonstrated that mononuclear BMCs transplantation was capable of increasing the levels of VEGF, FGF and Angiopoietin I expression in the infarction scar and its periphery, compared with the control group, P <0 05 Conclusion These findings indicate that the natural angiogenic processes leading to collateral development are extremely complex, since many kinds of bone marrow derived growth factors involved in the processes after mononuclear BMCs transplantation into infarction sites