College of Food Science and Engineering, Nanjing University of Finance and Economics/Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing 210023,China.
Engineering Research Center of Bio-process, Ministry of Education, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, Anhui, China.
J Agric Food Chem. 2023 Jun 7;71(22):8437-8447. doi: 10.1021/acs.jafc.3c00227. Epub 2023 May 24.
This study aimed to investigate the anti-inflammatory molecular activity of rapeseed napin-derived dipeptide Thr-Leu (TL) using Caco-2/RAW264.7 cell cocultures. This in vitro coculture intestinal inflammation model was used to assess the absorption, evolution, and anti-inflammatory effects of peptides. TL was absorbed by the intestinal epithelial cells with an apparent permeability of (2.48 ± 0.18) × 10 cm/s, primarily through the PepT1 pathway. TL treatment exerted anti-inflammatory and restorative effects on the impaired intestinal barrier function by enhancing the expression levels of occludin and ZO-1 in lipopolysaccharide (LPS)-induced Caco-2 cells. No significant change ( < 0.05) was detected in claudin-1 expression levels; however, the occludin expression levels were upregulated through the protein kinase C (PKC) signaling pathway. Compared with the LPS-induced group, TL (2.0 mM) reduced the levels of intracellular inflammation-related enzymes (iNOS: by 50.84%; COX-2: by 49.64%) on the coculture cell model. In addition, the interleukin (IL)-1β, IL-6, and TNF-α levels in RAW264.7 cells were significantly ( < 0.05) downregulated following TL treatment (2.0 mM) due to the suppression of the phosphorylation of the JNK-independent pathway on the basolateral side of the coculture cell model. These findings highlight the potential use of TL in functional foods or nutraceuticals to prevent intestinal inflammation.
本研究旨在利用 Caco-2/RAW264.7 细胞共培养物来研究油菜籽 napin 衍生二肽 Thr-Leu (TL) 的抗炎分子活性。该体外共培养肠道炎症模型用于评估肽的吸收、演变和抗炎作用。TL 通过肠上皮细胞的表观渗透率（2.48 ± 0.18）× 10 cm/s 被吸收，主要通过 PepT1 途径。TL 处理通过增强脂多糖 (LPS) 诱导的 Caco-2 细胞中 occludin 和 ZO-1 的表达水平，对受损的肠道屏障功能发挥抗炎和修复作用。Claudin-1 的表达水平没有明显变化（<0.05）；然而，occludin 的表达水平通过蛋白激酶 C (PKC) 信号通路被上调。与 LPS 诱导组相比，TL（2.0 mM）在共培养细胞模型中降低了细胞内炎症相关酶（iNOS：降低 50.84%；COX-2：降低 49.64%）的水平。此外，TL（2.0 mM）处理还显著降低了 RAW264.7 细胞中白细胞介素 (IL)-1β、IL-6 和 TNF-α的水平，这是由于共培养细胞模型基底外侧侧 JNK 非依赖性途径磷酸化的抑制所致。这些发现强调了 TL 在功能性食品或营养保健品中预防肠道炎症的潜在用途。
