The Fourth Affiliated Hospital of Zhejiang University School of Medicine, Yiwu, 322000, China.
Inflammation. 2015;38(3):1213-20. doi: 10.1007/s10753-014-0087-8.
The purpose of this study was to evaluate the effects of polydatin (PD) on cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expressions at protein and transcriptional levels, as well as the production of prostaglandin E2 (PGE2) and nitric oxide (NO) in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells. To elucidate the underlying mechanism responsible for these symptoms, we investigated the phosphorylation of mitogen-activated protein kinase (MAPK) pathway and nuclear factor-κB (NF-κB) expression. NO was analyzed with the Griess method. PGE2 was measured by enzyme-linked immunosorbent assay (ELISA). iNOS and COX-2 messenger RNA (mRNA) were identified by qPCR assay. iNOS, COX-2, NF-κB, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 protein expressions were detected with Western blot. The results revealed that PD effectively inhibited NO and PGE2 production, and it is not surprising that PD reduced iNOS and COX-2 expression at protein and transcriptional levels. Additionally, PD significantly ameliorated the activation of NF-κB and the phosphorylation of MAPKs in LPS-induced RAW 264.7 macrophages. These findings suggested that PD exerted potent anti-inflammatory activity in macrophages.
本研究旨在评估白藜芦醇（PD）对脂多糖（LPS）诱导的巨噬细胞 RAW 264.7 中环氧合酶-2（COX-2）和诱导型一氧化氮合酶（iNOS）蛋白和转录水平表达以及前列腺素 E2（PGE2）和一氧化氮（NO）产生的影响。为了阐明这些症状的潜在机制，我们研究了丝裂原活化蛋白激酶（MAPK）途径和核因子-κB（NF-κB）表达的磷酸化。采用格里斯法分析 NO。通过酶联免疫吸附试验（ELISA）测定 PGE2。通过 qPCR 检测 iNOS 和 COX-2 信使 RNA（mRNA）。用 Western blot 检测 iNOS、COX-2、NF-κB、细胞外信号调节激酶（ERK）、c-Jun N 末端激酶（JNK）和 p38 蛋白表达。结果表明，PD 能有效抑制 NO 和 PGE2 的产生，并不奇怪的是，PD 能降低 LPS 诱导的 RAW 264.7 巨噬细胞中 iNOS 和 COX-2 的蛋白和转录水平表达。此外，PD 显著改善了 LPS 诱导的 RAW 264.7 巨噬细胞中 NF-κB 的激活和 MAPKs 的磷酸化。这些发现表明 PD 在巨噬细胞中发挥了强大的抗炎活性。
