Alaaeldin Rania, Hassan Heba Ali, Abdel-Rahman Islam M, Mohyeldin Reham H, Youssef Nancy, Allam Ahmed E, Abdelwahab Sayed F, Zhao Qing-Li, Fathy Moustafa
Department of Biochemistry, Faculty of Pharmacy, Deraya University, Minia 61111, Egypt.
Department of Pharmacognosy, Faculty of Pharmacy, Sohag University, Sohag 82524, Egypt.
Curr Issues Mol Biol. 2022 Jul 2;44(7):2967-2981. doi: 10.3390/cimb44070205.
Inflammation is a critical defensive mechanism mainly arising due to the production of prostaglandins via cyclooxygenase enzymes. This study aimed to examine the anti-inflammatory activity of fatty acid glucoside (FAG), which is isolated from Ficus benghalensis against lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cytotoxic activity of the FAG on RAW 264.7 macrophages was evaluated with an MTT assay. The levels of PGE2 and NO and the activity of iNOS, COX-1, and COX-2 enzymes in LPS-stimulated RAW 264.7 cells were evaluated. The gene expression of IL-6, TNF-α, and PGE2 was investigated by qRT-PCR. The expression of epidermal growth factor receptor (EGFR), Akt, and PI3K proteins was examined using Western blotting analysis. Furthermore, molecular docking of the new FAG against EGFR was investigated. A non-cytotoxic concentration of FAG increased NO release and iNOS activity, inhibited COX-1 and COX-2 activities, and reduced PGE2 levels in LPS-stimulated RAW 264.7 cells. It diminished the expression of TNF-α, IL-6, PGE2, EGFR, Akt, and PI3K. Furthermore, the molecular docking study proposed the potential direct binding of FAG with EGFR with a high affinity. This study showed that FAG is a natural EGFR inhibitor, NO-releasing, and COX-inhibiting anti-inflammatory agent via EGFR/Akt/PI3K pathway inhibition.
炎症是一种关键的防御机制，主要是由于通过环氧化酶产生前列腺素而引发的。本研究旨在检测从孟加拉榕中分离出的脂肪酸糖苷（FAG）对脂多糖（LPS）刺激的RAW 264.7巨噬细胞的抗炎活性。采用MTT法评估FAG对RAW 264.7巨噬细胞的细胞毒性活性。评估了LPS刺激的RAW 264.7细胞中PGE2和NO的水平以及诱导型一氧化氮合酶（iNOS）、环氧化酶-1（COX-1）和环氧化酶-2（COX-2）的活性。通过qRT-PCR研究白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和PGE2的基因表达。使用蛋白质印迹分析检测表皮生长因子受体（EGFR）、蛋白激酶B（Akt）和磷脂酰肌醇-3激酶（PI3K）蛋白的表达。此外，还研究了新型FAG与EGFR的分子对接。FAG的非细胞毒性浓度增加了NO释放和iNOS活性，抑制了COX-1和COX-2活性，并降低了LPS刺激的RAW 264.7细胞中PGE2的水平。它减少了TNF-α、IL-6、PGE2、EGFR、Akt和PI3K的表达。此外，分子对接研究表明FAG与EGFR具有潜在的高亲和力直接结合。本研究表明，FAG是一种天然的EGFR抑制剂，通过抑制EGFR/Akt/PI3K途径具有释放NO和抑制COX的抗炎作用。
