Heterogeneous nuclear ribonucleoprotein E1 regulates protein disulphide isomerase translation in oxidized low-density lipoprotein-activated endothelial cells
Heterogeneous nuclear ribonucleoprotein E1 regulates protein disulphide isomerase translation in oxidized low-density lipoprotein-activated endothelial cells
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DOI：
10.1111/apha.12422
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发表时间：
2015-03-01
影响因子：
6.3
通讯作者：
Miao, J.
中科院分区：
文献类型：
--
作者：
Meng, N.;Peng, N.;Miao, J.
目的内皮源性蛋白二硫化物异构酶（PDI）是体内血栓形成所必需的。但是，如何控制氧化低密度脂蛋白（oxLDL）激活的血管内皮细胞（VECs）中PDI的过度产生还没有很好的理解。在这项研究中，我们试图回答这个问题，使用我们新发现的mTOC 1 3-苄基-5-（（2-硝基苯氧基）甲基）-二氢呋喃-2（3 H）-酮（3BDO），已被证明可以保护VECs.MethodsFirst，我们进行了蛋白质组学分析的oxLDL激活的血管VECs中存在或不存在的3BDO。其次，我们构建了异质核核糖核蛋白E1（hnRNP E1）的Ser 43突变体，并使用RNA-ChIP技术来研究hnRNP E1和PDI生产之间的关系。此外，我们研究了3BDO对oxLDL改变的Akt 1和Akt 2磷酸化的影响。最后，我们研究了3BDO对oxLDL改变的PDI蛋白水平在载脂蛋白E-/-小鼠晚期atherosclerosis.ResultsIn血管内皮细胞，oxLDL增加PDI蛋白水平，诱导hnRNP E1磷酸化Ser 43，抑制hnRNP E1结合到PDI 5非翻译区，并诱导磷酸化的Akt 2，但不是Akt 1。所有这些过程都被3BDO阻断。重要的是，hnRNP E1的Ser 43突变体抑制了oxLDL诱导的PDI蛋白水平的升高和hnRNP E1与PDI 5 UTR结合的降低。结论hnRNP E1是oxLDL激活的VECs中PDI翻译的一个新的调节因子，3BDO是一种有效的抑制PDI过度表达的药物。
AimsEndothelium-derived protein disulphide isomerase (PDI) is required for thrombus formation in vivo. But, how to control PDI overproduction in oxidized low-density lipoprotein (oxLDL)-activated vascular endothelial cells (VECs) is not well understood. In this study, we try to answer this question using our newly identified activator of mTOC1 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2 (3H)-one (3BDO) that has been shown to protect VECs.MethodsFirst, we performed a proteomics analysis on the oxLDL-activated vascular VECs in the presence or absence of 3BDO. Next, we constructed the heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) mutants at Ser43 and used the RNA-ChIP technique to investigate the relationship between hnRNP E1 and PDI production. Furthermore, we examined the effect of 3BDO on oxLDL-altered phosphorylation of Akt1 and Akt2. Finally, we studied the effect of 3BDO on oxLDL-altered PDI protein level in apolipoprotein E-/- mice with advanced atherosclerosis.ResultsIn VECs, oxLDL-increased PDI protein level, induced hnRNP E1 phosphorylation at Ser43, suppressed the binding of hnRNP E1 to PDI 5UTR and induced the phosphorylation of Akt2 but not Akt1. All ofthese processes were blocked by 3BDO. Importantly, Ser43 mutant of hnRNP E1 inhibited the increase of PDI protein level and the decrease of the binding of hnRNP E1 and PDI 5UTR induced by oxLDL. Furthermore, 3BDO suppressed oxLDL-induced PDI protein increase in the serum and plaque endothelium of apolipoprotein E-/- mice.ConclusionhnRNP E1 is a new regulator of PDI translation in oxLDL-activated VECs, and 3BDO is a powerful agent for controlling PDI overproduction.