Alcalá-Santiago Ángela, Toscano-Sánchez Rocío, Márquez-López José Carlos, González-Jurado José Antonio, Fernández-Pachón María-Soledad, García-Villanova Belén, Pedroche Justo, Rodríguez-Martín Noelia María
Department of Nutrition and Food Science, Faculty of Pharmacy, University of Granada, 18071 Granada, Spain.
Instituto de Investigación Biosanitaria ibs. GRANADA, 18012 Granada, Spain.
Int J Mol Sci. 2024 Nov 25;25(23):12628. doi: 10.3390/ijms252312628.
Vitamin D (VD), a crucial micronutrient, regulates bone health and immune responses. Recent studies suggest that VD may confer protective effects against chronic inflammatory diseases. Additionally, plant-based peptides can show biological activities. Furthermore, the supplementation of protein hydrolysates with VD could potentially enhance the bioactivity of peptides, leading to synergistic effects. In this study, THP-1 cells were exposed to low concentrations of Lipopolysaccharide (LPS) to induce inflammation, followed by treatment with vitamin D at different concentrations (10, 25, or 50 nM) or a chickpea protein hydrolysate ("H30BIO") supplemented with VD. The cytotoxicity of VD was evaluated using viability assay to confirm its safety. The cytokine secretion of TNF-α, IL-1β, and IL6 was assessed in the cell supernatant, and the gene expression of , , , , , , , , , , , , , and was quantified by qRT-PCR. Treatment with VD alone significantly decreased the expression of the pro-inflammatory genes and , as well as their corresponding cytokine levels in the supernatants. While gene expression remained unchanged, a reduction in its cytokine release was observed upon VD treatment. No dose-dependent effects were observed. Interestingly, the combination of VD with H30BIO led to an increase in expression and secretion in contrast with the LPS control, coupled with a decrease in levels. Additionally, genes such as , , , , , and exhibited notable modulation, suggesting that the combination treatment primarily downregulates NF-κB-related gene activity. This study demonstrates a synergistic interaction between VD and H30BIO, suggesting that this combination may enhance pathways involving TNF-α, potentially aiding in the resolution and modulation of inflammation through adaptive processes. These findings open new avenues for research into the therapeutic applications of enriched protein hydrolysates with VD to manage low-grade inflammatory-related conditions.
维生素D（VD）是一种关键的微量营养素，可调节骨骼健康和免疫反应。最近的研究表明，VD可能对慢性炎症性疾病具有保护作用。此外，植物源肽可表现出生物活性。此外，用VD补充蛋白质水解物可能会增强肽的生物活性，从而产生协同效应。在本研究中，THP-1细胞暴露于低浓度的脂多糖（LPS）以诱导炎症，随后用不同浓度（10、25或50 nM）的维生素D或补充了VD的鹰嘴豆蛋白水解物（“H30BIO”）进行处理。使用活力测定法评估VD的细胞毒性以确认其安全性。在细胞上清液中评估TNF-α、IL-1β和IL6的细胞因子分泌，并通过qRT-PCR对 、 、 、 、 、 、 、 、 、 、 、 、 和 的基因表达进行定量。单独用VD处理可显著降低促炎基因 和 的表达及其在上清液中的相应细胞因子水平。虽然 基因表达保持不变，但在VD处理后观察到其细胞因子释放减少。未观察到剂量依赖性效应。有趣的是，与LPS对照相比，VD与H30BIO的组合导致 表达和分泌增加，同时 水平降低。此外， 、 、 、 、 和 等基因表现出显著调节，表明联合治疗主要下调NF-κB相关基因活性。本研究证明了VD与H30BIO之间的协同相互作用，表明这种组合可能增强涉及TNF-α 的途径，潜在地通过适应性过程有助于炎症的消退和调节。这些发现为研究用VD富集蛋白质水解物在治疗与低度炎症相关病症中的应用开辟了新途径。
