Department of Pharmacological and Biomolecular Sciences "Rodolfo Paoletti" (DiSFeB), Università Degli Studi di Milano, 20133 Milan, Italy.
Naturalsalus S.r.l., Trezzano Rosa, 20060 Milan, Italy.
Nutrients. 2024 Aug 17;16(16):2743. doi: 10.3390/nu16162743.
Sulforaphane is considered the bioactive metabolite of glucoraphanin after dietary consumption of broccoli sprouts. Although both molecules pass through the gut lumen to the large intestine in stable form, their biological impact on the first intestinal tract is poorly described. In celiac patients, the function of the small intestine is affected by celiac disease (CD), whose severe outcomes are controlled by gluten-free dietary protocols. Nevertheless, pathological signs of inflammation and oxidative stress may persist. The aim of this study was to compare the biological activity of sulforaphane with its precursor glucoraphanin in a cellular model of gliadin-induced inflammation. Human intestinal epithelial cells (CaCo-2) were stimulated with a pro-inflammatory combination of cytokines (IFN-γ, IL-1β) and in-vitro-digested gliadin, while oxidative stress was induced by HO. LC-MS/MS analysis confirmed that sulforaphane from broccoli sprouts was stable after simulated gastrointestinal digestion. It inhibited the release of all chemokines selected as inflammatory read-outs, with a more potent effect against MCP-1 (IC = 7.81 µM). On the contrary, glucoraphanin (50 µM) was inactive. The molecules were unable to counteract the oxidative damage to DNA (γ-H2AX) and catalase levels; however, the activity of NF-κB and Nrf-2 was modulated by both molecules. The impact on epithelial permeability (TEER) was also evaluated in a Transwell model. In the context of a pro-inflammatory combination including gliadin, TEER values were recovered by neither sulforaphane nor glucoraphanin. Conversely, in the context of co-culture with activated macrophages (THP-1), sulforaphane inhibited the release of MCP-1 (IC = 20.60 µM) and IL-1β (IC = 1.50 µM) only, but both molecules restored epithelial integrity at 50 µM. Our work suggests that glucoraphanin should not merely be considered as just an inert precursor at the small intestine level, thus suggesting a potential interest in the framework of CD. Its biological activity might imply, at least in part, molecular mechanisms different from sulforaphane.
萝卜硫素被认为是食用西兰花芽后萝卜硫苷的生物活性代谢物。尽管这两种分子都以稳定的形式穿过肠腔到达大肠，但它们对第一肠道的生物学影响描述得很差。在乳糜泻患者中，小肠的功能受到乳糜泻（CD）的影响，其严重后果受无麸质饮食方案的控制。然而，炎症和氧化应激的病理迹象可能仍然存在。本研究旨在比较萝卜硫素与其前体萝卜硫苷在麦胶蛋白诱导炎症的细胞模型中的生物学活性。人肠上皮细胞（CaCo-2）用细胞因子（IFN-γ、IL-1β）和体外消化的麦胶蛋白的促炎混合物刺激，同时用 HO 诱导氧化应激。LC-MS/MS 分析证实，来自西兰花芽的萝卜硫素在模拟胃肠道消化后是稳定的。它抑制了所有作为炎症读出物选择的趋化因子的释放，对 MCP-1 的抑制作用更强（IC = 7.81 µM）。相反，萝卜硫苷（50 µM）没有活性。这些分子不能对抗 DNA（γ-H2AX）和过氧化氢酶水平的氧化损伤；然而，NF-κB 和 Nrf-2 的活性被这两种分子调节。还在 Transwell 模型中评估了对上皮通透性（TEER）的影响。在包括麦胶蛋白的促炎混合物的情况下，无论是萝卜硫素还是萝卜硫苷都不能恢复 TEER 值。相反，在与激活的巨噬细胞（THP-1）共培养的情况下，萝卜硫素仅抑制 MCP-1（IC = 20.60 µM）和 IL-1β（IC = 1.50 µM）的释放，但两种分子都能在 50 µM 时恢复上皮完整性。我们的工作表明，萝卜硫苷不应仅仅被认为是小肠水平上的一种惰性前体，因此在 CD 的框架内具有潜在的意义。它的生物学活性可能至少部分暗示了与萝卜硫素不同的分子机制。
