Dietary flavones 6,3´,4´-trihydroxyflavone (6,3´,4´-HOFL) and 7,3´,4´-trihydroxyflavone (7,3´,4´-HOFL) showed preliminary antioxidant and anti-inflammatory activities in a two-dimensional (2D) cell culture model. However, their action mechanisms remain unclear, and the anti-inflammatory activities have not been studied in a reliable three-dimensional (3D) cell model. Therefore, in the current study, the antioxidant potency was examined by their scavenging ability of cellular reactive oxygen species. Anti-inflammatory activities were examined via their inhibitory effects on inflammatory mediators in vitro on 2D and 3D macrophage models, and their mechanisms were determined through transcriptome. In the 3D macrophages, two flavones were less bioactive than they were in 2D macrophages, but they both significantly suppressed the overexpression of proinflammatory mediators in two cell models. The divergent position of the hydroxyl group on the A ring resulted in activity differences. Compared to 6,3´,4´-HOFL, 7,3´,4´-HOFL showed lower activity on NO, IL-1β suppression, and c-Src binding (IC50: 12.0 and 20.9 µM) but higher ROS-scavenging capacity (IC50: 3.20 and 2.71 µM) and less cytotoxicity. In addition to the IL-17 and TNF pathways of 6,3´,4´-HOFL, 7,3´,4´-HOFL also exerted anti-inflammatory activity through JAK-STAT, as indicated by the RNA-sequencing results. Two flavones showed prominent antioxidant and anti-inflammatory activities on 2D and 3D models.

中文翻译：

---

6,3',4'- 和 7,3',4'-三羟基黄酮对 2D 和 3D RAW264.7 模型的抗氧化和抗炎作用。

膳食黄酮 6,3',4'-三羟基黄酮 (6,3',4'-HOFL) 和 7,3',4'-三羟基黄酮 (7,3',4'-HOFL) 具有初步的抗氧化和抗炎作用二维 (2D) 细胞培养模型中的活动。然而，它们的作用机制仍不清楚，并且尚未在可靠的三维 (3D) 细胞模型中研究其抗炎活性。因此，在目前的研究中，抗氧化效力是通过它们清除细胞活性氧的能力来检验的。通过它们在 2D 和 3D 巨噬细胞模型上对体外炎症介质的抑制作用来检查抗炎活性，并通过转录组确定它们的机制。在 3D 巨噬细胞中，两种黄酮的生物活性低于 2D 巨噬细胞，但它们都显着抑制了两种细胞模型中促炎介质的过度表达。A环上羟基的不同位置导致活性差异。与 6,3',4'-HOFL 相比，7,3',4'-HOFL 对 NO、IL-1β 抑制和 c-Src 结合的活性较低（IC50：12.0 和 20.9 µM），但 ROS 清除能力较高容量（IC50：3.20 和 2.71 µM）和较低的细胞毒性。如 RNA 测序结果所示，除了 6,3',4'-HOFL 的 IL-17 和 TNF 通路外，7,3',4'-HOFL 还通过 JAK-STAT 发挥抗炎活性。两种黄酮在 2D 和 3D 模型上显示出显着的抗氧化和抗炎活性。4´-HOFL 对 NO、IL-1β 抑制和 c-Src 结合显示出较低的活性（IC50：12.0 和 20.9 µM），但具有更高的 ROS 清除能力（IC50：3.20 和 2.71 µM）和较低的细胞毒性。如 RNA 测序结果所示，除了 6,3',4'-HOFL 的 IL-17 和 TNF 通路外，7,3',4'-HOFL 还通过 JAK-STAT 发挥抗炎活性。两种黄酮在 2D 和 3D 模型上显示出显着的抗氧化和抗炎活性。4´-HOFL 对 NO、IL-1β 抑制和 c-Src 结合显示出较低的活性（IC50：12.0 和 20.9 µM），但具有更高的 ROS 清除能力（IC50：3.20 和 2.71 µM）和较低的细胞毒性。如 RNA 测序结果所示，除了 6,3',4'-HOFL 的 IL-17 和 TNF 通路外，7,3',4'-HOFL 还通过 JAK-STAT 发挥抗炎活性。两种黄酮在 2D 和 3D 模型上显示出显着的抗氧化和抗炎活性。