This study aimed to identify alternative anti-inflammatory compounds that modulate the activity of a relevant transcription factor, CCAAT/enhancer binding protein delta (C/EBPδ). C/EBPδ is a master regulator of inflammatory responses in macrophages (Mϕ) and is mainly regulated at the level of CEBPD gene transcription initiation. To screen for CEBPD-modulating compounds, we generated a THP-1-derived reporter cell line stably expressing secreted alkaline phosphatase (SEAP) under control of the defined CEBPD promoter (CEBPD::SEAP). A high-throughput screening of LOPAC^®1280 and ENZO^®774 libraries on LPS- and IFN-γ-activated THP-1 reporter Mϕ identified four epigenetically active hits: two bromodomain and extraterminal domain (BET) inhibitors, I-BET151 and Ro 11-1464, as well as two histone deacetylase (HDAC) inhibitors, SAHA and TSA. All four hits markedly and reproducibly upregulated SEAP secretion and CEBPD::SEAP mRNA expression, confirming screening assay reliability. Whereas BET inhibitors also upregulated the mRNA expression of the endogenous CEBPD, HDAC inhibitors completely abolished it. All hits displayed anti-inflammatory activity through the suppression of IL-6 and CCL2 gene expression. However, I-BET151 and HDAC inhibitors simultaneously upregulated the mRNA expression of pro-inflammatory IL-1ß. The modulation of CEBPD gene expression shown in this study contributes to our understanding of inflammatory responses in Mϕ and may offer an approach to therapy for inflammation-driven disorders.

中文翻译：

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在THP-1衍生的记者巨噬细胞中对CEBPD调节化合物进行高通量筛选，可确定抗炎性HDAC和BET抑制剂

这项研究旨在鉴定可调节相关转录因子CCAAT /增强子结合蛋白δ（C /EBPδ）活性的替代抗炎化合物。C /EBPδ是巨噬细胞（Mϕ）中炎症反应的主要调节剂，主要在CEBPD基因转录起始水平上调节。为了筛选调节CEBPD的化合物，我们在定义的CEBPD启动子（CEBPD :: SEAP）的控制下，生成了稳定表达分泌型碱性磷酸酶（SEAP）的THP-1衍生的报告细胞系。^LOPAC®1280和^ENZO®774的高通量筛选LPS和IFN-γ激活的THP-1报道分子Mϕ的文库鉴定了四个表观遗传活性的命中点：两种溴结构域和末端结构域（BET）抑制剂I-BET151和Ro 11-1464，以及两种组蛋白脱乙酰基酶（HDAC）抑制剂，SAHA和TSA。所有四个命中均显着且可复制地上调了SEAP分泌和CEBPD :: SEAP mRNA表达，从而证实了筛选测定的可靠性。BET抑制剂也上调了内源性CEBPD的mRNA表达，而HDAC抑制剂则完全消除了它。所有命中都通过抑制IL-6和CCL2基因表达而表现出抗炎活性。但是，I-BET151和HDAC抑制剂同时上调了促炎药的mRNA表达IL-1ß。这项研究中显示的CEBPD基因表达的调节有助于我们了解Mϕ中的炎症反应，并可能为炎症引起的疾病的治疗提供一种方法。