脯氨酰羟化酶结构域蛋白 2 沉默增强了移植的脂肪源性干细胞在梗死心肌中的存活和旁分泌功能。
Prolyl hydroxylase domain protein 2 silencing enhances the survival and paracrine function of transplanted adipose-derived stem cells in infarcted myocardium.
机构信息
Department of Cardiology, Daping Hospital.
出版信息
Circ Res. 2013 Jul 19;113(3):288-300. doi: 10.1161/CIRCRESAHA.113.300929. Epub 2013 May 21.
RATIONALE
Transplantation of stem cells into damaged hearts has had modest success as a treatment for ischemic heart disease. One of the limitations is the poor stem cell survival in the diseased microenvironment. Prolyl hydroxylase domain protein 2 (PHD2) is a cellular oxygen sensor that regulates 2 key transcription factors involved in cell survival and inflammation: hypoxia-inducible factor and nuclear factor-κB.
OBJECTIVE
We studied whether and how PHD2 silencing in human adipose-derived stem cells (ADSCs) enhances their cardioprotective effects after transplantation into infarcted hearts.
METHODS AND RESULTS
ADSCs were transduced with lentiviral short hairpin RNA against prolyl hydroxylase domain protein 2 (shPHD2) to silence PHD2. ADSCs, with or without shPHD2, were transplanted after myocardial infarction in mice. ADSCs reduced cardiomyocyte apoptosis, fibrosis, and infarct size and improved cardiac function. shPHD2-ADSCs exerted significantly more protection. PHD2 silencing induced greater ADSC survival, which was abolished by short hairpin RNA against hypoxia-inducible factor-1α. Conditioned medium from shPHD2-ADSCs decreased cardiomyocyte apoptosis. Insulin-like growth factor-1 (IGF-1) levels were significantly higher in the conditioned medium of shPHD2-ADSCs versus ADSCs, and depletion of IGF-1 attenuated the cardioprotective effects of shPHD2-ADSC-conditioned medium. Nuclear factor-κB activation was induced by shPHD2 to induce IGF-1 secretion via binding to IGF-1 gene promoter.
CONCLUSIONS
PHD2 silencing promotes ADSCs survival in infarcted hearts and enhances their paracrine function to protect cardiomyocytes. The prosurvival effect of shPHD2 on ADSCs is hypoxia-inducible factor-1α dependent, and the enhanced paracrine function of shPHD2-ADSCs is associated with nuclear factor-κB-mediated IGF-1 upregulation. PHD2 silencing in stem cells may be a novel strategy for enhancing the effectiveness of stem cell therapy after myocardial infarction.
理由
将干细胞移植到受损的心脏中已被证明是治疗缺血性心脏病的一种有效方法。其中一个限制因素是干细胞在病变微环境中的存活率较低。脯氨酰羟化酶结构域蛋白 2（PHD2）是一种细胞氧传感器，可调节涉及细胞存活和炎症的 2 个关键转录因子：缺氧诱导因子和核因子-κB。
目的
我们研究了沉默人脂肪来源干细胞（ADSCs）中的 PHD2 是否以及如何增强其在移植到梗死心脏后的心脏保护作用。
方法和结果
ADSCs 被慢病毒短发夹 RNA（shRNA）转导以沉默 PHD2。在小鼠心肌梗死后，将 ADSC 转染或未转染 shPHD2。ADSC 减少了心肌细胞凋亡、纤维化和梗死面积，并改善了心脏功能。shPHD2-ADSC 发挥了更显著的保护作用。PHD2 沉默诱导了更多的 ADSC 存活，而这种存活被针对缺氧诱导因子-1α的短发夹 RNA 所消除。shPHD2-ADSC 的条件培养基降低了心肌细胞凋亡。胰岛素样生长因子-1（IGF-1）水平在 shPHD2-ADSC 的条件培养基中明显高于 ADSC，而 IGF-1 的耗竭减弱了 shPHD2-ADSC 条件培养基的心脏保护作用。核因子-κB 的激活诱导 IGF-1 的分泌，通过与 IGF-1 基因启动子结合。
结论
PHD2 沉默促进了梗死心脏中 ADSC 的存活，并增强了其旁分泌功能以保护心肌细胞。shPHD2 对 ADSC 的促生存作用依赖于缺氧诱导因子-1α，而 shPHD2-ADSC 增强的旁分泌功能与核因子-κB 介导的 IGF-1 上调有关。干细胞中 PHD2 的沉默可能是增强心肌梗死后干细胞治疗效果的一种新策略。
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