Department of Food and Life Sciences, College of Biomedical Science & Engineering, Inje University, Gimhae 621-749, Republic of Korea.
Oxid Med Cell Longev. 2013;2013:409321. doi: 10.1155/2013/409321. Epub 2013 Oct 23.
In the present study, the anti-inflammatory effect and underlying mechanisms of wild grape seeds procyanidins (WGP) were examined using lipopolysaccharide- (LPS-) stimulated RAW 264.7 cells. We used nitric oxide (NO) and prostaglandin E2 (PGE2) and reactive oxygen species (ROS) assays to examine inhibitory effect of WGP and further investigated the mechanisms of WGP suppressed LPS-mediated genes and upstream expression by Western blot and confocal microscopy analysis. Our data indicate that WGP significantly reduced NO, PGE2, and ROS production and also inhibited the expression of proinflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions. Consistently, WGP significantly reduced LPS-stimulated expression of proinflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin- (IL-) 1 β . Moreover, WGP prevented nuclear translocation of nuclear factor- κ B (NF κ B) p65 subunit by reducing inhibitory κ B- α (I κ B α) and NF κ B phosphorylation. Furthermore, we found that WGP inhibited LPS-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK). Taken together, our results demonstrated that WGP exerts potent anti-inflammatory activity through the inhibition of iNOS and COX-2 by regulating NF κ B and p38 MAPK pathway.
在本研究中,使用脂多糖(LPS)刺激的 RAW 264.7 细胞来研究野生葡萄籽原花青素(WGP)的抗炎作用及其潜在机制。我们使用一氧化氮(NO)和前列腺素 E2(PGE2)和活性氧(ROS)测定法来检查 WGP 的抑制作用,并通过 Western blot 和共聚焦显微镜分析进一步研究了 WGP 抑制 LPS 介导的基因和上游表达的机制。我们的数据表明,WGP 可显著减少 NO、PGE2 和 ROS 的产生,并抑制诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2)蛋白表达等促炎介质的表达。一致地,WGP 还显著降低了 LPS 刺激的促炎细胞因子如肿瘤坏死因子-α(TNF-α)和白细胞介素-(IL-)1β的表达。此外,WGP 通过减少 IκBα和 NF-κB 磷酸化来阻止核因子-κB(NF-κB)p65 亚基的核易位。此外,我们发现 WGP 抑制了 LPS 诱导的 p38 丝裂原活化蛋白激酶(MAPK)的磷酸化。总之,我们的结果表明,WGP 通过调节 NF-κB 和 p38 MAPK 途径抑制 iNOS 和 COX-2 的表达发挥强大的抗炎作用。