School of Food Engineering and Biotechnology, Hanshan Normal University, Chaozhou 521041, China.
Shandong Center for Drug and Food Evaluation & Certification, Jinan 250014, China.
J Zhejiang Univ Sci B. 2019;20(10):803-815. doi: 10.1631/jzus.B1800631.
Activated nuclear factor-κB (NF-κB) plays an important role in the development of cardiovascular disease (CVD) through its regulated genes and microRNAs (miRNAs). However, the gene regulation profile remains unclear. In this study, primary mouse vascular endothelial cells (pMVECs) were employed to detect CVD-related NF-κB-regulated genes and miRNAs. Genechip assay identified 77 NF-κB-regulated genes, including 45 upregulated and 32 downregulated genes, in tumor necrosis factor α (TNFα)-treated pMVECs. Ten of these genes were also found to be regulated by NF-κB in TNFα-treated HeLa cells. Quantitative real-time PCR (qRT-PCR) assay confirmed the up-regulation of Egr1, Tnf, and Btg2 by NF-κB in the TNFα-treated pMVECs. The functional annotation revealed that many NF-κB-regulated genes identified in pMVECs were clustered into classical NF-κB-involved biological processes. Genechip assay also identified 26 NF-κB-regulated miRNAs, of which 21 were upregulated and 5 downregulated, in the TNFα-treated pMVECs. Further analysis showed that nine of the identified genes are regulated by seven of these miRNAs. Finally, among the identified NF-κB-regulated genes and miRNAs, 5 genes and 12 miRNAs were associated with CVD by miRWalk and genetic association database analysis. Taken together, these findings show an intricate gene regulation network raised by NF-κB in TNFα-treated pMVECs. The network provides new insights for understanding the molecular mechanism underlying the progression of CVD.
激活的核因子-κB（NF-κB）通过其调控的基因和 microRNAs（miRNAs）在心血管疾病（CVD）的发展中发挥重要作用。然而，基因调控谱尚不清楚。在这项研究中，我们使用原代小鼠血管内皮细胞（pMVECs）来检测 TNFα 处理的 pMVECs 中与 CVD 相关的 NF-κB 调控基因和 miRNAs。基因芯片分析鉴定出 77 个 NF-κB 调控基因，包括 45 个上调和 32 个下调基因，在 TNFα 处理的 pMVECs 中。其中 10 个基因也被发现在 TNFα 处理的 HeLa 细胞中受到 NF-κB 的调控。定量实时 PCR（qRT-PCR）检测证实 NF-κB 在 TNFα 处理的 pMVECs 中上调了 Egr1、Tnf 和 Btg2 的表达。功能注释表明，在 pMVECs 中鉴定出的许多 NF-κB 调控基因被聚类到经典的 NF-κB 参与的生物学过程中。基因芯片分析还鉴定出 26 个 NF-κB 调控的 miRNAs，其中 21 个上调，5 个下调，在 TNFα 处理的 pMVECs 中。进一步分析表明，鉴定出的基因中有 9 个受到其中 7 个 miRNAs 的调控。最后，在鉴定出的 NF-κB 调控基因和 miRNAs 中，miRWalk 和遗传关联数据库分析显示，有 5 个基因和 12 个 miRNAs 与 CVD 相关。总之，这些发现表明 NF-κB 在 TNFα 处理的 pMVECs 中引起了复杂的基因调控网络。该网络为理解 CVD 进展的分子机制提供了新的见解。
