Comparative effectiveness of 4 natural and chemical activators of Nrf2 on inflammation, oxidative stress, macrophage polarization, and bactericidal activity in an in vitro macrophage infection model
Comparative effectiveness of 4 natural and chemical activators of Nrf2 on inflammation, oxidative stress, macrophage polarization, and bactericidal activity in an in vitro macrophage infection model
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DOI：
10.1371/journal.pone.0234484
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发表时间：
2020-06-08
期刊：
影响因子：
3.7
通讯作者：
Deramaudt, Therese B.
中科院分区：
文献类型：
--
作者：
Ali, Malika;Bonay, Marcel;Deramaudt, Therese B.
炎症在先天免疫系统对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个具有潜在或已证实的抗炎和/或抗微生物特性的化合物，在我们的细菌感染THP-1来源的巨噬细胞的体外模型上进行测试。我们首先比较了萝卜硫素(SFN)、汉黄素(WG)、奥替普拉兹(OTZ)和富马酸二甲酯(DMF)诱导核因子红系相关因子2(Nrf2)的能力，Nrf2是抗氧化、抗炎反应途径的关键调节因子。接下来，我们对所选择的4种化合物的抗氧化和抗炎效果进行了比较评估。分别用THP-1诱导的巨噬细胞和内毒素刺激的巨噬细胞处理THP-1诱导的巨噬细胞，用RT-qPCR方法检测IL-1β、IL-6和TNF-α基因的表达水平。此外，用每种化合物处理经典激活的M1巨噬细胞，检测编码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，检测各化合物对革兰氏阴性杆菌和革兰氏阳性金黄色葡萄球菌在THP-1来源的巨噬细胞和PBMC来源的巨噬细胞内细菌存活的影响。我们的数据证实了三七总皂甙、WG和DMF对内毒素刺激的THP-1来源的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来源的巨噬细胞降低了M1标记基因的表达水平，而SFN或DMF处理上调了M2标记基因MRC1的表达水平。这种M1标记基因表达的减少可能与SFN或DMF处理的巨噬细胞内金黄色葡萄球菌载量的减少有关。有趣的是，在SFN处理的THP-1来源的巨噬细胞中，大肠杆菌的细胞内存活率增加，这在PBMC来源的巨噬细胞中没有观察到。相反，OTZ表现出促氧化和促炎特性，并影响THP-1来源的巨噬细胞中大肠杆菌的细胞内存活。总之，我们在治疗炎症和细菌感染方面提供了新的潜在治疗选择。
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-1 beta, IL-6, and TNF-alpha were quantified by RT-qPCR. Moreover, expression levels of genes coding for M1 (IL-23, CCR7, IL-1 beta, IL-6, and TNF-alpha) and M2 (PPAR gamma, 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.