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ER stress regulates alkaline phosphatase gene expression in vascular smooth muscle cells via an ATF4-dependent mechanism

机译:内质网应激通过ATF4依赖性机制调节血管平滑肌细胞中碱性磷酸酶基因的表达

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Abstract ObjectiveVascular calcification is the deposition of hydroxyapatite crystals in the blood vessel wall. Osteogenic differentiation of vascular smooth muscle cells (VSMCs) plays a key role in this process. Increased expression of alkaline phosphatase (ALP) occurs in some in vitro?models of VSMC calcification and is thought to be crucial for mineralization, however, little is known about the transcriptional regulation of ALP in VSMCs. Recently, ALP upregulation was shown to coincide with endoplasmic reticulum (ER) stress-mediated vascular calcification, specifically with expression of the transcription factor ATF4. As no direct links between ALP expression and ER stress have previously been demonstrated in VSMCs, the aim of this study was to investigate whether ATF4 interacts directly with the ALP promoter.ResultsThe present study shows that ALP mRNA and activity were significantly increased by ER stress treatment of human primary VSMCs in vitro and that this was ATF4-dependent. Bioinformatics analysis predicted two ATF4 binding sites in ER-stress responsive regions of the ALP promoter (??3631 to ??2048?bp from the first intron). However, we found that ATF4 does not bind within this fragment of the ALP promoter region.
机译:摘要目的血管钙化是羟基磷灰石晶体在血管壁上的沉积。血管平滑肌细胞(VSMC)的成骨分化在此过程中起关键作用。在某些VSMC钙化的体外模型中,碱性磷酸酶(ALP)的表达增加,被认为对矿化至关重要,但是,关于VSMC中ALP的转录调控了解甚少。最近,显示ALP上调与内质网(ER)应激介导的血管钙化相吻合,特别是与转录因子ATF4的表达相吻合。由于以前尚未在VSMC中证实ALP表达与内质网应激之间存在直接联系,因此本研究的目的是研究ATF4是否与ALP启动子直接相互作用。结果本研究表明,内质网应激处理可显着提高ALP mRNA和活性。人原代VSMC的体外培养,并且这是ATF4依赖性的。生物信息学分析预测了ALP启动子的ER应激反应区域中的两个ATF4结合位点(从第一个内含子开始约3631到2048 bp)。但是,我们发现ATF4不在ALP启动子区域的此片段内结合。

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