Upregulation of VEGF by small activating RNA and its implications in preeclampsia
Upregulation of VEGF by small activating RNA and its implications in preeclampsia
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小激活 RNA 上调 VEGF 及其对先兆子痫的影响
DOI：
10.1016/j.placenta.2016.08.088
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发表时间：
2016-10-01
期刊：
影响因子：
3.8
通讯作者：
Yu, Jun
中科院分区：
文献类型：
--
作者：
Guo, Xijiao;Feng, Ling;Yu, Jun
前言：先兆子痫是一种严重的妊娠并发症，主要是由于血管扩张不足和螺旋动脉重塑。VEGF是血管生成的主要因子，是调节胎盘血管生成过程所必需的。因此，降低VEGF在妊娠期高血压也可能导致低灌注和随后的缺氧的胎儿在高血压pregnances.Methods：本研究旨在阐明VEGF的作用机制在先兆子痫。使用小活化RNA（saRNA）上调人滋养层细胞（HTR-8/SVneo）中的VEGF表达。采用实时荧光定量PCR和Western blot检测VEGF的表达水平，流式细胞仪检测转染效率。通过伤口划痕试验分析细胞迁移。通过测定NO代谢产物检测NO分泌。Western blot分析eNOS表达。结果：转染saRNA后，VEGF表达明显增加（P均< 0.05），而转染后VEGF表达无明显变化（P> 0.05）。在HTR-8/SVneo细胞中，saRNA显著增加NO分泌和eNOS表达（分别为p = 0.0003和0.032）。HTR-8/SVneo细胞的迁移能力和管形成功能被saRNA增强（分别为p = 0.024和0.013）。TNIF-alpha抑制VEGF下游eNOS-NO通路的活性以及细胞迁移和小管形成，同时加强VEGF的表达减弱了TNF-α诱导的所有损伤。结论：利用RNA激活策略来增加内源性VEGF的表达可能是一种新兴的和有效的治疗先兆子痫的方法。(C)2016爱思唯尔有限公司版权所有。
Introduction: Preeclampsia is a severe pregnancy complication mostly due to inadequate vascular dilation and remodeling of spiral arteries. VEGF, the major factor for angiogenesis, is necessary for modulating angiogenic processes in the placenta. Hence reduction of VEGF in gestational hypertension may also lead to hypoperfusion and subsequent hypoxia of the fetus in hypertensive pregnancy.Methods: This study aimed at elucidating the mechanism of action of VEGF in preeclampsia. Small activating RNAs (saRNA) were used to upregulate VEGF expression in human trophoblast cells (HTR-8/SVneo). The VEGF expression level was analyzed by real-time quantitative PCR and western blot, while its transfection efficiency was measured by flow cytometer assay. Cell migration was analyzed by a wound scratch assay. NO secretion was detected by determining NO metabolites. eNOS expression was analyzed by western blot. Tube formation function of cells was then analyzed by matrigel migration assay.Results: VEGF expression significantly increased after saRNA transfection (all p < 0.05). NO secretion and eNOS expression significantly increased by saRNA in HTR-8/SVneo cells (p = 0.0003 and 0.032 respectively). The migration ability and tube formation function of HTR-8/SVneo cells were enhanced by saRNA (p = 0.024 and 0.013 respectively). TNIF-alpha inhibited VEGF-downstream eNOS-NO pathway activity as well as cell migration and tubulogenesis, while enforcing the expression of VEGF attenuated all the insults induced by TNF-alpha.Conclusions: Utilizing an RNA activation strategy to increase endogenous VEGF expression could be an emerging and effective approach for the treatment of preeclampsia. (C) 2016 Elsevier Ltd. All rights reserved.