心肌细胞靶向 HIF-1α 基因治疗抑制大鼠心肌细胞凋亡和心脏移植血管病变。
Cardiomyocyte-targeted HIF-1alpha gene therapy inhibits cardiomyocyte apoptosis and cardiac allograft vasculopathy in the rat.
机构信息
Cardiopulmonary Research Group, Transplantation Laboratory, Haartman Institute, University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland.
出版信息
J Heart Lung Transplant. 2010 Sep;29(9):1058-66. doi: 10.1016/j.healun.2010.05.021. Epub 2010 Jun 26.
BACKGROUND
Hypoxia-inducible factor-1 (HIF-1), a key transcription factor in hypoxia, affects a wide range of adaptive cell functions. We examined the kinetics of endogenous HIF-1alpha during acute and chronic rejection, and the effect of exogenous HIF-1alpha in chronically rejecting rat cardiac allografts.
METHODS
Heterotopic cardiac transplantations were performed between major MHC-mismatched Dark Agouti and Wistar-Furth rats. Cyclosporine A (CsA) was used to prevent acute rejection in the chronic rejection model. The effect of HIF-1alpha overexpression was investigated by adeno-assocated virus 2 (AAV2)-mediated gene transfer of a constitutively stabilized form of mouse HIF-1alpha (AAV-HIF-1alpha). The analysis of allografts was based on histology, immunohistochemistry and quantitative reverse transcript-polymerase chain reaction (RT-PCR).
RESULTS
Acute and chronic rejection significantly induced HIF-1alpha mRNA in rat cardiac allografts when compared with syngeneic controls. Immunohistochemistry localized significantly increased HIF-1alpha immunoreactivity to vascular smooth muscle cells, vascular endothelial cells, post-capillary venules and graft-infiltrating mononuclear inflammatory cells of the allograft, whereas expression in cardiomyocytes remained unchanged. Regression analysis revealed a linear correlation between the progression of cardiac allograft vasculopathy (CAV) and HIF-1alpha immunoreactivity in post-capillary venules and graft-infiltrating mononuclear inflammatory cells in chronically rejecting rat cardiac allografts. AAV-HIF-1alpha enhanced cardiomyocyte HIF-1alpha production and significantly reduced cardiomyocyte apoptosis and the development of CAV in chronically rejecting rat cardiac allografts.
CONCLUSIONS
We found that acute and chronic rejection increased HIF-1alpha mRNA and protein levels in rat cardiac allografts. On the other hand, cardiomyocyte-targeted HIF-1alpha gene transfer inhibited cardiomyocyte apoptosis and the development of CAV, suggesting a novel therapeutic strategy for HIF-1alpha in cardiac allografts.
背景
缺氧诱导因子-1（HIF-1）是缺氧条件下的关键转录因子，影响广泛的适应性细胞功能。我们研究了内源性 HIF-1α 在急性和慢性排斥反应中的动力学，以及外源性 HIF-1α 在慢性排斥反应大鼠心脏同种异体移植物中的作用。
方法
在主要 MHC 错配的 Dark Agouti 和 Wistar-Furth 大鼠之间进行异位心脏移植。环孢素 A（CsA）用于预防慢性排斥反应模型中的急性排斥反应。通过腺相关病毒 2（AAV2）介导的稳定形式的小鼠 HIF-1α（AAV-HIF-1α）的基因转移来研究 HIF-1α 过表达的影响。同种异体移植物的分析基于组织学、免疫组织化学和定量逆转录聚合酶链反应（RT-PCR）。
结果
与同基因对照相比，急性和慢性排斥反应显著诱导大鼠心脏同种异体移植物中的 HIF-1α mRNA。免疫组织化学将显著增加的 HIF-1α 免疫反应性定位到血管平滑肌细胞、血管内皮细胞、毛细血管后小静脉和同种异体移植物中的浸润单核炎性细胞，而心肌细胞中的表达保持不变。回归分析显示，慢性排斥反应大鼠心脏同种异体移植物中毛细血管后小静脉和浸润单核炎性细胞的心脏同种异体移植物血管病变（CAV）进展与 HIF-1α 免疫反应性之间存在线性相关性。AAV-HIF-1α 增强了心肌细胞 HIF-1α 的产生，并显著减少了慢性排斥反应大鼠心脏同种异体移植物中的心肌细胞凋亡和 CAV 的发展。
结论
我们发现急性和慢性排斥反应增加了大鼠心脏同种异体移植物中的 HIF-1α mRNA 和蛋白水平。另一方面，心肌细胞靶向 HIF-1α 基因转移抑制了心肌细胞凋亡和 CAV 的发展，表明 HIF-1α 在心脏同种异体移植物中具有新的治疗策略。
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