Hwang Da Young, Ki Dae-Won, Choi Dae-Cheol, Yun Bong-Sik, Kim Yoon Hee
Department of Food and Nutrition, College of Engineering, Daegu University, Gyeongsan-si 38453, Republic of Korea.
Division of Biotechnology, Jeonbuk National University, Iksan-si 54596, Republic of Korea.
Biomedicines. 2025 Jun 12;13(6):1445. doi: 10.3390/biomedicines13061445.
Pestalotic acid A (PAA), a polyketide derived from , an endophyte of the Japanese holly (), is known to exhibit known antimicrobial activity, but its anti-inflammatory properties remain uncharacterized. This study aimed to investigate the anti-inflammatory effects of PAA in lipopolysaccharide (LPS)-stimulated murine macrophages, RAW264.7 cells. PAA was isolated from endophytes of and its structure was confirmed. RAW264.7 macrophages were treated with 0-50 μM of PAA in the presence of 100 ng/mL LPS. Cell viability was assessed by MTS assay; nitric oxide (NO) production was measured via Griess reagent; interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF) were quantified by enzyme-linked immunosorbent assay. Protein expression of inducible NO synthase (iNOS), nuclear factor (NF)-κB p65 phosphorylation, and related signaling proteins was evaluated by Western blot analysis and immunofluorescence staining. PAA significantly increased macrophage viability and dose-dependently inhibited the release of NO by alleviating the protein expression of iNOS in LPS-treated RAW264.7 cells. Furthermore, PAA suppressed the release of IL-6, IL-1β, and TNF induced by LPS. Western blot and immunofluorescence results also indicated that PAA blocked the p65 subunit phosphorylation of NF-κB, which is one of the underlying mechanisms of the anti-inflammatory action of pestalotic acid A. PAA exerts potent anti-inflammatory effects in LPS-stimulated macrophages via inhibition of the NF-κB pathway, highlighting its potential as a natural therapeutic agent for inflammatory diseases.
盘多毛孢酸A（PAA）是一种从日本冬青（Ilex serrata）的内生菌 Pestalotiopsis sp. 中提取的聚酮化合物，已知具有抗菌活性，但其抗炎特性尚未得到表征。本研究旨在探讨PAA对脂多糖（LPS）刺激的小鼠巨噬细胞RAW264.7细胞的抗炎作用。从Pestalotiopsis sp. 的内生菌中分离出PAA并确认了其结构。在存在100 ng/mL LPS的情况下，用0-50 μM的PAA处理RAW264.7巨噬细胞。通过MTS法评估细胞活力；通过格里斯试剂测量一氧化氮（NO）的产生；通过酶联免疫吸附测定法定量白细胞介素（IL）-6、IL-1β和肿瘤坏死因子（TNF）。通过蛋白质印迹分析和免疫荧光染色评估诱导型一氧化氮合酶（iNOS）的蛋白质表达、核因子（NF）-κB p65磷酸化及相关信号蛋白。PAA显著提高了巨噬细胞活力，并通过减轻LPS处理的RAW264.7细胞中iNOS的蛋白质表达，剂量依赖性地抑制了NO的释放。此外，PAA抑制了LPS诱导的IL-6、IL-1β和TNF的释放。蛋白质印迹和免疫荧光结果还表明，PAA阻断了NF-κB的p65亚基磷酸化，这是盘多毛孢酸A抗炎作用的潜在机制之一。PAA通过抑制NF-κB途径在LPS刺激的巨噬细胞中发挥强大的抗炎作用，突出了其作为炎症性疾病天然治疗剂的潜力。
