UVSQ, INSERM END-ICAP, Université Paris-Saclay, Versailles, France.
Service de Physiologie-Explorations Fonctionnelles, Hôpital Ambroise Paré, APHP, Boulogne, France.
PLoS One. 2020 Jun 8;15(6):e0234484. doi: 10.1371/journal.pone.0234484. eCollection 2020.
Inflammation plays a crucial role in the defense response of the innate immune system against pathogen infection. In this study, we selected 4 compounds for their potential or proven anti-inflammatory and/or anti-microbial properties to test on our in vitro model of bacteria-infected THP-1-derived macrophages. We first compared the capacity of sulforaphane (SFN), wogonin (WG), oltipraz (OTZ), and dimethyl fumarate (DMF) to induce the nuclear factor erythroid 2-related factor 2 (Nrf2), a key regulator of the antioxidant, anti-inflammatory response pathways. Next, we performed a comparative evaluation of the antioxidant and anti-inflammatory efficacies of the 4 selected compounds. THP-1-derived macrophages and LPS-stimulated macrophages were treated with each compound and expression levels of genes coding for inflammatory cytokines IL-1β, IL-6, and TNF-α were quantified by RT-qPCR. Moreover, expression levels of genes coding for M1 (IL-23, CCR7, IL-1β, IL-6, and TNF-α) and M2 (PPARγ, MRC1, CCL22, and IL-10) markers were determined in classically-activated M1 macrophages treated with each compound. Finally, the effects of each compound on the intracellular bacterial survival of gram-negative E. coli and gram-positive S. aureus in THP-1-derived macrophages and PBMC-derived macrophages were examined. Our data confirmed the anti-inflammatory and antioxidant effects of SFN, WG, and DMF on LPS-stimulated THP-1-derived macrophages. In addition, SFN or WG treatment of classically-activated THP-1-derived macrophages reduced expression levels of M1 marker genes, while SFN or DMF treatment upregulated the M2 marker gene MRC1. This decrease in expression of M1 marker genes may be correlated with the decrease in intracellular S. aureus load in SFN- or DMF-treated macrophages. Interestingly, an increase in intracellular survival of E. coli in SFN-treated THP-1-derived macrophages that was not observed in PBMC-derived macrophages. Conversely, OTZ exhibited pro-oxidant and proinflammatory properties, and affected intracellular survival of E. coli in THP-1-derived macrophages. Altogether, we provide new potential therapeutic alternatives in treating inflammation and bacterial infection.
炎症在先天免疫系统抵御病原体感染的防御反应中起着至关重要的作用。在这项研究中，我们选择了 4 种化合物，因为它们具有潜在的或已被证实的抗炎和/或抗微生物特性，以便在我们体外细菌感染的 THP-1 衍生巨噬细胞模型上进行测试。我们首先比较了萝卜硫素（SFN）、白杨素（WG）、奥替普拉（OTZ）和富马酸二甲酯（DMF）诱导核因子红细胞 2 相关因子 2（Nrf2）的能力，Nrf2 是抗氧化、抗炎反应途径的关键调节剂。接下来，我们对这 4 种选定化合物的抗氧化和抗炎功效进行了比较评估。用每种化合物处理 THP-1 衍生的巨噬细胞和 LPS 刺激的巨噬细胞，并通过 RT-qPCR 定量测定编码炎症细胞因子 IL-1β、IL-6 和 TNF-α的基因的表达水平。此外，还测定了用每种化合物处理的经典激活的 M1 巨噬细胞中编码 M1（IL-23、CCR7、IL-1β、IL-6 和 TNF-α）和 M2（PPARγ、MRC1、CCL22 和 IL-10）标志物的基因的表达水平。最后，检查了每种化合物对 THP-1 衍生巨噬细胞和 PBMC 衍生巨噬细胞中革兰氏阴性大肠杆菌和革兰氏阳性金黄色葡萄球菌的细胞内存活的影响。我们的数据证实了 SFN、WG 和 DMF 对 LPS 刺激的 THP-1 衍生巨噬细胞的抗炎和抗氧化作用。此外，SFN 或 WG 处理经典激活的 THP-1 衍生巨噬细胞降低了 M1 标志物基因的表达水平，而 SFN 或 DMF 处理则上调了 M2 标志物基因 MRC1。SFN 或 DMF 处理的巨噬细胞中 M1 标志物基因表达的降低可能与 SFN 或 DMF 处理的巨噬细胞中金黄色葡萄球菌载量的降低有关。有趣的是，SFN 处理的 THP-1 衍生巨噬细胞中大肠杆菌的细胞内存活增加，而在 PBMC 衍生的巨噬细胞中则没有观察到这种情况。相反，OTZ 表现出促氧化和促炎特性，并影响 THP-1 衍生巨噬细胞中大肠杆菌的细胞内存活。总的来说，我们为治疗炎症和细菌感染提供了新的潜在治疗选择。
