College of Animal Science and Technology, Jilin Agricultural University, Changchun, 130062, People's Republic of China.
Inflammation. 2014 Feb;37(1):235-46. doi: 10.1007/s10753-013-9734-8.
The aim of this study was to investigate the protective effects of cepharanthine (CEP) on inflammation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in vitro and a LPS-induced lung injury model in vivo. RAW264.7 cells were treated with various concentrations of CEP for 1 h followed by incubation with or without 1 μg/ml LPS for 18 h. TNF-α, IL-6, and IL-1β in the supernatants were measured by ELISA. Nuclear factor-κB (NF-κB) and mitogen-activated protein kinase pathways were analyzed by Western blot. Mice were randomly divided into control group, LPS group, CEP + LPS group, and dexamethasone + LPS group. A male BALB/c mouse model of acute lung injury was induced by LPS. Bronchoalveolar lavage fluid was collected for inflammatory cell count and cytokine assays. Histopathologic examination was performed on mice that were not subjected to bronchoalveolar lavage fluid collection. CEP dose-dependently inhibited the release of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. Significantly, CEP dose-dependently suppressed NF-κB activation, IκBα degradation, and phosphorylation of ERK, JNK, and p38 induced by LPS. In vivo, it was also observed that CEP attenuated lung histopathologic changes and down-regulated the level of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, in the mouse acute lung injury model. These results suggest that CEP potentially decreases inflammation in vitro and in vivo and might be a therapeutic agent against inflammatory diseases.
本研究旨在探讨苦参碱（CEP）对体外脂多糖（LPS）刺激的 RAW264.7 细胞炎症以及体内 LPS 诱导的肺损伤模型的保护作用。用不同浓度的 CEP 处理 RAW264.7 细胞 1 小时，然后用或不用 1μg/ml LPS 孵育 18 小时。通过 ELISA 测定上清液中 TNF-α、IL-6 和 IL-1β 的含量。用 Western blot 分析核因子-κB（NF-κB）和丝裂原活化蛋白激酶途径。将小鼠随机分为对照组、LPS 组、CEP+LPS 组和地塞米松+LPS 组。用 LPS 诱导雄性 BALB/c 小鼠急性肺损伤模型。收集支气管肺泡灌洗液进行炎症细胞计数和细胞因子检测。对未进行支气管肺泡灌洗的小鼠进行组织病理学检查。CEP 呈剂量依赖性抑制 LPS 刺激的 RAW264.7 细胞中 TNF-α、IL-6 和 IL-1β 的释放。重要的是，CEP 呈剂量依赖性抑制 LPS 诱导的 NF-κB 激活、IκBα降解以及 ERK、JNK 和 p38 的磷酸化。在体内，还观察到 CEP 减轻了肺组织病理学变化，并下调了小鼠急性肺损伤模型中促炎细胞因子 TNF-α、IL-1β 和 IL-6 的水平。这些结果表明 CEP 可能在体外和体内减轻炎症，并且可能是一种治疗炎症性疾病的药物。
