School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Shenzhen Affliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Int Immunopharmacol. 2018 Mar;56:90-97. doi: 10.1016/j.intimp.2018.01.023. Epub 2018 Jan 23.
Sepsis is a life-threatening disease. Inflammation is a major concomitant symptom of sepsis Chrysophanol, an anthraquinone derivative isolated from the rhizomes of rheumpalmatum, has been reported to have a protective effect against lipopolysaccharide(LPS)-induced inflammation. However, the underlying molecular mechanisms are not well understood. The aim of this study was to explore the effect and mechanism of chrysophanol on lipopolysaccharide (LPS)-induced anti-inflammatory effect of RAW264.7 cells and its involved potential mechanism. The mRNA and protein expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and inducible nitric oxide synthase (iNOS), nuclear factor kappa B (NF-κB) and PPAR-γ were measured by qRT-PCR and western blotting, the production of TNF-α, IL-1β was evaluated by ELISA. Then, the phosphorylation of NF-κB p65 was also detected by western blotting. And NF-κB p65 promoter activity was analyzed by the Dual-Luciferase reporter assay system as well. Meanwhile, PPAR-γ inhibitor GW9662 was performed to knockdown PPAR-γ expression in cells. Our data revealed that LPS induced the up-regulation of TNF-α, IL-1β, iNOS and NF-κB p65, the down-regulation of PPAR-γ were substantially suppressed by chrysophanol in RAW264.7 cells. Furthermore, our data also figured out that these effects of chrysophanol were largely abrogated by PPAR-γ inhibitor GW9662. Taken together, our results indicated that LPS-induced inflammation was potently compromised by chrysophanol very likely through the PPAR-γ-dependent inactivation of NF-κB in RAW264.7 cells.
脓毒症是一种危及生命的疾病。炎症是脓毒症的主要伴随症状。从大黄根茎中分离得到的蒽醌衍生物大黄素已被报道具有对抗脂多糖（LPS）诱导的炎症的保护作用。然而，其潜在的分子机制尚不清楚。本研究旨在探讨大黄素对 LPS 诱导的 RAW264.7 细胞抗炎作用及其潜在机制。通过 qRT-PCR 和 Western blot 检测肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β 和诱导型一氧化氮合酶（iNOS）、核因子 kappa B（NF-κB）和过氧化物酶体增殖物激活受体-γ（PPAR-γ）的 mRNA 和蛋白表达，通过 ELISA 评估 TNF-α、IL-1β 的产生。然后，通过 Western blot 检测 NF-κB p65 的磷酸化。并通过双荧光素酶报告基因检测系统分析 NF-κB p65 启动子活性。同时，在细胞中用 PPAR-γ 抑制剂 GW9662 进行 PPAR-γ 表达的敲低。我们的数据表明，LPS 诱导 TNF-α、IL-1β、iNOS 和 NF-κB p65 的上调，大黄素显著抑制 RAW264.7 细胞中 PPAR-γ 的下调。此外，我们的数据还表明，大黄素的这些作用很大程度上被 PPAR-γ 抑制剂 GW9662 所阻断。综上所述，我们的研究结果表明，大黄素可能通过 PPAR-γ 依赖性 NF-κB 失活来抑制 LPS 诱导的 RAW264.7 细胞炎症。
