血管紧张素Ⅱ1型受体在脂多糖诱导RAW264.7巨噬细胞促炎性细胞因子产生中的作用  

Role of angiotensin Ⅱ type 1 receptor in the production of pro-inflammatory cytokines induced by lipopolysaccharide in RAW264.7 microphage

作　　者：郭峰[1] 陈旭林[1] 王飞[1] 王永杰[1] 孙业祥[1] 

机构地区：[1]安徽医科大学第一附属医院烧伤科,合肥230022

出　　处：《中华损伤与修复杂志（电子版）》2011年第2期19-22,共4页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：国家自然科学基金(30872687)

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。Objective To investigate the role of angiotensin Ⅱ type 1 receptor in the production of pro-inflammatory eytokines induced by lipopolysaccharide （LPS） in RAW264.7 microphage and characterize the mechanism. Methods RAW264.7 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-KB） and activator protein 1 （AP-1）. Results Compared 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 LIPS 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-KB and AP-1 to 1.43-fold （P 〈 0.01 ） and 1.90-fold （P 〈 0. O1 ） of control group. But compared with LPS group, the expression of TNF-α and IL-1β was inhibited significantly by the preineubation 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 maerophages induced by LPS via the activation of transcription factor NF-κB and

关 键 词：受体 血管紧张素 1型 脂多糖类 巨噬细胞 细胞因子类 核因子-κB 

分 类 号：R363[医药卫生—病理学]