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首页> 外文期刊>American Journal of Physiology >Embryonic stem cell differentiation into smooth muscle cells is mediated by Nox4-produced H2O2.
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Embryonic stem cell differentiation into smooth muscle cells is mediated by Nox4-produced H2O2.

机译：Nox4产生的H2O2介导胚胎干细胞向平滑肌细胞的分化。

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摘要

NADPH oxidase (Nox4) produces reactive oxygen species (ROS) that are important for vascular smooth muscle cell (SMC) behavior, but the potential impact of Nox4 in stem cell differentiation is unknown. When mouse embryonic stem (ES) cells were plated on collagen IV-coated dishes/flasks, a panel of SMC-specific genes was significantly and consistently upregulated. Nox4 expression was markedly correlated with such a gene induction as confirmed by real-time PCR, immunofluorescence, and Western blot analysis. Overexpression of Nox4 specifically resulted in increased SMC marker production, whereas knockdown of Nox4 induced a decrease. Furthermore, SMC-specific transcription factors, including serum response factor (SRF) and myocardin were activated by Nox4 gene expression. Moreover, Nox4 was demonstrated to drive SMC differentiation through generation of H(2)O(2). Confocal microscopy analysis indicates that SRF was translocated into the nucleus during SMC differentiation in which SRF was phosphorylated. Additionally, autosecreted transforming growth factor (TGF)-beta(1) activated Nox4 and promoted SMC differentiation. Interestingly, cell lines generated from stem cells by Nox4 transfection and G418 selection displayed a characteristic of mature SMCs, including expression of SMC markers and cells with contractile function. Thus we demonstrate for the first time that Nox4 is crucial for SMC differentiation from ES cells, and enforced Nox4 expression can maintain differentiation status and functional features of stem cell-derived SMCs, highlighting its impact on vessel formation in vivo and vascular tissue engineering in the future.
机译：NADPH氧化酶（Nox4）产生的活性氧（ROS）对血管平滑肌细胞（SMC）行为很重要，但Nox4对干细胞分化的潜在影响尚不清楚。当将小鼠胚胎干（ES）细胞铺在胶原蛋白IV包被的培养皿/培养皿上时，一组SMC特异性基因显着且持续上调。通过实时PCR，免疫荧光和蛋白质印迹分析证实，Nox4表达与这种基因诱导显着相关。 Nox4的过表达专门导致SMC标记物产生增加，而敲低Nox4则导致其减少。此外，SMC特异性转录因子（包括血清反应因子（SRF）和心肌素）被Nox4基因表达激活。此外，Nox4被证明可以通过生成H（2）O（2）驱动SMC分化。共聚焦显微镜分析表明，在SRF被磷酸化的SMC分化过程中，SRF易位到细胞核中。此外，自动分泌的转化生长因子（TGF）-beta（1）激活Nox4并促进SMC分化。有趣的是，通过Nox4转染和G418选择从干细胞生成的细胞系显示出成熟SMC的特征，包括SMC标记物的表达和具有收缩功能的细胞。因此，我们首次证明Nox4对于从ES细胞分化SMC至关重要，并且强制执行的Nox4表达可以维持干细胞衍生SMC的分化状态和功能特征，从而突出了其对体内血管形成和血管组织工程的影响。未来。

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