穿心莲内酯对活化巨噬细胞细胞外信号调节激酶1/2信号转导通路的抑制作用  

Andrographolide inhibits extracellular signal-regulated kinase 1/2 signaling pathway in activated macrophages

作　　者：覃林花[1] 吕礁[1] 孔玲[2] 施云兴[1] 李永平[1] 周国中[1] 郑智武[1] 李琳[1] 季兴英[1] 

机构地区：[1]中国人民解放军第四一一医院消化内科,上海200081 [2]中国科学院上海生命科学院健康科学研究中心,上海200025

出　　处：《中西医结合学报》2011年第6期632-637,共6页Journal of Chinese Integrative Medicine

摘　　要：目的：观察穿心莲内酯对脂多糖激活的小鼠腹腔巨噬细胞细胞外信号调节激酶1/2（extracellularsignal-regulated kinase1/2,ERK1/2）信号转导通路及肿瘤坏死因子α（tumor necrosisfactor-α,TNF-α）的影响。方法：以穿心莲内酯作用于脂多糖活化的小鼠腹腔巨噬细胞后,细胞计数试剂盒8测定穿心莲内酯对巨噬细胞的细胞毒作用,蛋白质印迹法检测巨噬细胞ERK1/2、p38和JNK蛋白磷酸化水平以及IκBα蛋白表达水平,逆转录聚合酶链反应法检测巨噬细胞TNF-αmRNA的表达水平,酶联免疫吸附测定法检测巨噬细胞培养上清液中TNF-α蛋白的表达水平。结果：1~100μg/mL穿心莲内酯对脂多糖活化的小鼠腹腔巨噬细胞无细胞毒作用;穿心莲内酯能抑制脂多糖活化小鼠腹腔巨噬细胞的ERK1/2磷酸化,随着穿心莲内酯浓度增加,抑制作用增强（P〈0.01）;1~25μg/mL穿心莲内酯不能抑制脂多糖活化小鼠腹腔巨噬细胞的p38和JNK磷酸化,对脂多糖引起的IκBα降解也无影响。12μg/mL穿心莲内酯和20μmol/LPD98059（ERK1/2信号转导通路抑制剂）能降低巨噬细胞TNF-αmRNA的表达和其上清液中TNF-α的分泌水平（P〈0.01）。结论：穿心莲内酯能阻断ERK1/2信号转导通路,并在抑制TNF-α的表达中可能起作用。Objective：To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2（ERK1/2）signaling pathway and tumor necrosis factor-α（TNF-α）expression in lipopolysaccharide（LPS）-activated macrophages.Methods：LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide.Cytotoxicity of andrographolide was detected by cell counting kit-8.The macrophages were lysed,and then expressions of phosphorylated ERK1/2,JNK and p38 and nuclear factor-κB inhibitor（IκBα）protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction.Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay.Results：Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages.Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages,which was concentration-dependent（P0.01）.Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages.In activated macrophages,TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059（inhibitor of ERK1/2 signaling pathway）at both mRNA expression and protein secretion levels.Conclusion：In LPS-activated macrophages,andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.

关 键 词：穿心莲内酯 细胞外信号调节MAP激酶类 丝裂原激活蛋白激酶类 肿瘤坏死因子α 巨噬细胞活化 

分 类 号：R285.5[医药卫生—中药学]