Demirbağ Burcu, Yıldırım Metin, Çimentepe Mehmet, Necip Adem, Ünver Hakan, Tiftik Eyüp Naci
Department of Stem Cell and Regenerative Medical, Institute of Health Sciences, Mersin University, Mersin, Türkiye.
Department of Biochemistry, Faculty of Pharmacy, Harran University, Sanliurfa, Türkiye.
Biochem Biophys Res Commun. 2025 Jun 20;766:151872. doi: 10.1016/j.bbrc.2025.151872. Epub 2025 Apr 22.
The rising incidence of pathogenic microorganisms underscores the urgent need to develop innovative anti-inflammatory agents. This study aims to investigate the anti-inflammatory and antibacterial mechanisms of Schiff base (SB) derived from salicylaldehyde (SA) compounds in stimulated and unstimulated macrophage cell line (RAW264.7) cells. We synthesized and characterized SB derived from SA using H NMR and C NMR spectroscopy. Anti-bacterial and computational insights of compounds were carried out. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was conducted to evaluate the toxicity profile of compounds on both lipopolysaccharide (LPS)-stimulated and unstimulated RAW264.7 cells. Additionally, the levels of nuclear factor kappa B (NFκB), interleukin (IL-6), tumor necrosis factor alpha (TNF-α), and reactive oxygen species (ROS) in the cells were measured using a microplate reader. The compound SA-SB-1 (the half maximal inhibitory concentration (IC: 39.58 μM) demonstrated strong inhibitory activity compared to SA-SB-2 (IC: 55.64 μM). SA-SB-1 exhibited greater inhibitory effects on the expression of NFκB, IL-6, TNF-α, and ROS in LPS-stimulated RAW264.7 cells than SA-SB-2. SA-SB-1 and SA-SB-2 exhibited a minimum inhibitory concentration (MIC) value of 12 μg/mL against E. faecalis and 250 μg/mL against S. aureus, P. aeruginosa, and E. coli. The binding affinities of SA-SB-1 and SA-SB-2 were studied against various proteins, including those with PDB IDs: 2AZ5, 4ZS7, 2RAM, 3G7B, 6QXS, and 2UV0. The highest binding affinity of SA-SB-2 was found to be -9.329 kcal/mol for 2UV0. These findings are promising for the potential development of new anti-inflammatory drugs.
致病微生物发病率的上升凸显了开发创新抗炎药物的迫切需求。本研究旨在探讨源自水杨醛（SA）化合物的席夫碱（SB）在刺激和未刺激的巨噬细胞系（RAW264.7）细胞中的抗炎和抗菌机制。我们使用氢核磁共振（¹H NMR）和碳核磁共振（¹³C NMR）光谱对源自SA的SB进行了合成和表征。对化合物进行了抗菌和计算分析。进行了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验，以评估化合物对脂多糖（LPS）刺激和未刺激的RAW264.7细胞的毒性。此外，使用酶标仪测量细胞中核因子κB（NFκB）、白细胞介素（IL-6）、肿瘤坏死因子α（TNF-α）和活性氧（ROS）的水平。与SA-SB-2（半数抑制浓度（IC₅₀）：55.64 μM）相比，化合物SA-SB-1（IC₅₀：39.58 μM）表现出较强的抑制活性。SA-SB-1对LPS刺激的RAW264.7细胞中NFκB、IL-6、TNF-α和ROS表达的抑制作用比SA-SB-2更大。SA-SB-1和SA-SB-2对粪肠球菌的最低抑菌浓度（MIC）值为12 μg/mL，对金黄色葡萄球菌、铜绿假单胞菌和大肠杆菌的MIC值为250 μg/mL。研究了SA-SB-1和SA-SB-2对各种蛋白质的结合亲和力，包括蛋白质数据银行（PDB）ID为：2AZ5、4ZS7、2RAM、3G7B、6QXS和2UV0的蛋白质。发现SA-SB-2对2UV0的最高结合亲和力为-9.329 kcal/mol。这些发现为新型抗炎药物的潜在开发带来了希望。
