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作 者:马春芳[1,2] 王寅[1,2] 沈爱娟[1,2] 蔡宛如[1,2]
机构地区:[1]浙江中医药大学附属第二医院 [2]浙江省新华医院,浙江杭州31005
出 处:《中华中医药学刊》2015年第6期1320-1323,共4页Chinese Archives of Traditional Chinese Medicine
基 金:国家自然科学基金项目(81273678)
摘 要:目的:研究大黄素对脂多糖(Lipopolysacchride,LPS)诱导后巨噬细胞RAW264.7释放炎症因子的影响及其分子机制。方法:将细胞分为6组,空白组,LPS组和不同浓度(1μM,5μM,10μM,25μM)大黄素+LPS组,不同浓度的大黄素预处理细胞2 h后,再加入1μg/m L LPS刺激细胞,用ELISA法检测细胞上清液中的TNF-α,IL-1β,IL-6蛋白水平,用RT-PCR法检测炎症因子mRNA表达,用Western-Blot检测核转录因子(NF-κB p65)的磷酸化。结果:LPS刺激后RAW264.7巨噬细胞炎症因子TNF-α,IL-1β,IL-6的表达和分泌明显高于空白组,而给予大黄素的LPS组表达和分泌上述细胞因子明显降低,且呈剂量依赖性。LPS刺激后的细胞NF-κB p65磷酸化明显增强,而大黄素则能抑制NF-κB p65的磷酸化,且呈剂量依赖性。结论:大黄素能明显抑制巨噬细胞释放炎症因子,降低mRNA的表达,其作用机制主要通过抑制NF-κB p65磷酸化的信号通路。Objective: To investigate the anti- inflammatory effect of emodin on cytokine production by lipopolysaccharide- induced macrophage RAW264. 7 cell and its molecular mechanism. Methods: The macrophage cells RAW264. 7were randomly divided into six groups: blank group,LPS group,different concentrations of emodin( 1 μM,5 μM,10 μM,25 μM) + LPS groups. All groups cells were pretreated with different concentrations for 2 h,and then followed by 1 μg /m L LPS. The protein levels of TNF- α,IL- 1β,IL- 6 were detected by ELISA assay and its mRNA levels were measured by RT- PCR and NF- κB p65 phosphorylation was detected by Western- blot. Result: The expression and secretion of TNF- α,IL- 1β,IL- 6 in LPS group were increased significantly than those of blank groups,while emodin suppressed its protein and mRNA level in a dose- dependent manner. The NF- κB p65 phosphorylation was stronger than that of blank group while emodin could block the phosphorylation in a dose- dependent manner. Conclusion: Emodin can inhibit the mRNA expression and extracellar secretion of inflammatory cytokines production and its molecular mechanism is due to the inhibition of NF- κB p65 phosphorylation.
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