【摘要】 目的探讨血管紧张素Ⅱ1型受体在脂多糖(LPS)诱导巨噬细胞促炎性细胞因子产生中的作用和机制。方法按照随机数字表法将小鼠单核巨噬细胞株(RAW264.7)细胞分为空白对照组、ZD7155组、LPS组和LPS+ZD7155组。酶联免疫吸附试验法(ELISA)测定各组细胞培养上清液中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的含量,逆转录聚合酶链式反应(RT-PCR)检测RAW264.7细胞内TNF-α和IL-1β mRNA表达,凝胶电泳迁移检测法(EMSA)检测RAW264.7细胞内核因子-κB(NF-κB)和活化蛋白-1(AP-1)活性的变化。结果和空白对照组相比,ZD7155组培养上清液中的TNF-α和IL-1β含量差异没有统计学意义(P>0.05),但是LPS组细胞培养上清液中的TNF-α和IL-1β含量均显著高于空白对照组(均P<0.01)和ZD7155组(均P<0.05)。LPS组细胞内的TNF-α和IL-1βmRNA表达是空白对照组的2.19倍(P<0.01)和1.77倍(P<0.01),细胞内NF-κB和AP-1活性是空白对照组的1.43倍(P<0.01)和1.90倍(P<0.01)。而LPS+ZD7155组上清液中的TNF-α和IL-1β含量较LPS组下降(均P<0.05),细胞内TNF-α和IL-1βmRNA表达较LPS组下降了34.7%(P<0.01)和49.72%(P<0.01),同时细胞内NF-κB活性和AP-1活性较LPS组下降了46.15%(P<0.05)和48.42%(P<0.05)。结论血管紧张素Ⅱ1型受体通过活化转录因子NF-κB和AP-1,参与了LPS诱导巨噬细胞促炎性细胞因子TNF-α和IL-1β的产生和释放。更多还原

【Abstract】 Objective To investigate the role of angiotensin Ⅱ type 1 receptor in the production of pro-inflammatory cytokines induced by lipopolysaccharide(LPS) in RAW264.7 microphage and characterize the mechanism.MethodsRAW264.7 macrophages were randomly divided into four groups:control group,ZD7155 group,LPS group and ZD7155 + LPS group.The protein and mRNA expressions of tumor necrosis factor α(TNF-α) and interleukin-1β(IL-1β) in the cells were determined by enzyme-linked immunosorbent assay(ELISA) and reverse transcription polymerase chain reaction(RT-PCR) respectively.Electrophoretic mobility shift assay(EMSA) was preformed to determine the activities of nuclear factor kappaB(NF-κB) and activator protein 1(AP-1).ResultsCompared with control group,supernatant TNF-α and IL-1β of ZD7155 group were not found statistically significant difference(P>0.05).However,LPS stimulation increased the levels of TNF-α and IL-1β in the supernatant which were significantly higher than control group(P<0.01) and ZD7155 group(P<0.05).However,administration of LPS not only enhanced the mRNA expressions of TNF-α and IL-1β to 2.19-fold(P<0.01) and 1.77-fold(P<0.01) of control group,but also elevated the activities of NF-κB and AP-1 to 1.43-fold(P<0.01) and 1.90-fold(P<0.01) of control group.But compared with LPS group,the expression of TNF-α and IL-1β was inhibited significantly by the preincubation with ZD7155,intracellular TNF-α and IL-1β mRNA expression decreased by 34.7%(P<0.01) and 49.72%(P<0.01).Furthermore,compared with LPS group,the intracellular NF-κB and AP-1 activity of ZD7155+LPS group decreased by 46.15%(P<0.05) and 48.42%(P<0.05).Conclusion Angiotensin Ⅱ 1 receptor mediates the production and release of pro-inflammatory cytokines TNF-α and IL-1β in macrophages induced by LPS via the activation of transcription factor NF-κB and AP-1.更多还原