腺病毒介导的肝细胞生长因子诱导的坏死性凋亡激活内源性C-Kit+心脏干细胞并促进梗死老年心脏中的心肌细胞增殖和血管生成。

Necroptosis Induced by Ad-HGF Activates Endogenous C-Kit+ Cardiac Stem Cells and Promotes Cardiomyocyte Proliferation and Angiogenesis in the Infarcted Aged Heart.

作者信息

Liu Jiabao, Wu Peng, Wang Hao, Wang Yunle, Du Yingqiang, Cheng Weili, Xu Zhihui, Zhou Ningtian, Wang Liansheng, Yang Zhijian

机构信息

Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

出版信息

Cell Physiol Biochem. 2016;40(5):847-860. doi: 10.1159/000453144. Epub 2016 Dec 7.

DOI:10.1159/000453144

PMID:27941320

Abstract

BACKGROUND/AIMS: The discovery of c-kit+ cardiac stem cells (CSCs) provided us with new therapeutic targets to repair the damaged heart. However, the precise mechanisms regulating CSC proliferation and differentiation in the aged heart remained elusive. Necroptosis, a type of regulated cell death, has recently been shown to occur following myocardial infarction (MI); however, its effect on c-kit+ CSCs remains unknown. We investigated the effects of hepatocyte growth factor (HGF) and necroptosis on the proliferation and differentiation of endogenous c-kit+ CSCs in aged rat hearts following MI.

METHODS

The c-kit+ CSCs and HGF/p-Met expression levels in neonatal, adult and aged rats were compared using immunofluorescence and Western blotting. Immediately after MI, adenovirus carrying the HGF gene (Ad-HGF) was injected into the left ventricular wall surrounding the infarct areas of the aged rat heart. The proliferation and differentiation of the endogenous c-kit+ CSCs were studied using immunofluorescence. The signalling pathways were analysed via Western blotting and ELISA.

RESULTS

HGF/p-Met expression levels and c-kit+ CSC abundance gradually decreased with age. Ad-HGF promoted c-kit+ CSC differentiation into precursor cells of cardiomyocyte, endothelial and smooth muscle cell lineages and enhanced cardiomyocyte proliferation and angiogenesis in aged rats; these effects were reversed by the inhibition of necroptosis. Ad-HGF administration induced necroptosis by increasing the expression of receptor interacting protein kinase (RIP) 1 and receptor interacting protein kinase (RIP) 3 proteins in the infarcted heart. Moreover, Ad-HGF-induced necroptosis increased high-mobility group box 1 protein (HMGB1) levels and enhanced the abundance of c-kit+ cells in the bone marrow, which may partly account for the beneficial effect of necroptosis on the c-kit+ CSCs.

CONCLUSION

Ad-HGF-induced necroptosis facilitated aged heart repair after MI by promoting c-kit+ CSC proliferation and differentiation. These findings may lead to the development of new methods for the treatment of ischaemic heart disease in aged populations.

摘要

背景/目的：c-kit+心脏干细胞（CSCs）的发现为我们修复受损心脏提供了新的治疗靶点。然而，衰老心脏中调节CSC增殖和分化的精确机制仍不清楚。坏死性凋亡是一种程序性细胞死亡，最近研究表明其在心肌梗死（MI）后发生；然而，其对c-kit+CSCs的影响尚不清楚。我们研究了肝细胞生长因子（HGF）和坏死性凋亡对MI后老年大鼠心脏内源性c-kit+CSCs增殖和分化的影响。

方法

采用免疫荧光和蛋白质印迹法比较新生、成年和老年大鼠中c-kit+CSCs和HGF/p-Met的表达水平。MI后立即将携带HGF基因的腺病毒（Ad-HGF）注射到老年大鼠心脏梗死区域周围的左心室壁。采用免疫荧光法研究内源性c-kit+CSCs的增殖和分化。通过蛋白质印迹法和酶联免疫吸附测定法分析信号通路。

结果

HGF/p-Met表达水平和c-kit+CSC丰度随年龄增长逐渐降低。Ad-HGF促进老年大鼠c-kit+CSCs分化为心肌细胞、内皮细胞和平滑肌细胞系的前体细胞，并增强心肌细胞增殖和血管生成；抑制坏死性凋亡可逆转这些作用。给予Ad-HGF通过增加梗死心脏中受体相互作用蛋白激酶（RIP）1和受体相互作用蛋白激酶（RIP）3蛋白的表达诱导坏死性凋亡。此外，Ad-HGF诱导的坏死性凋亡增加了高迁移率族蛋白B1（HMGB1）水平，并增加了骨髓中c-kit+细胞的丰度，这可能部分解释了坏死性凋亡对c-kit+CSCs的有益作用。