脂肪细胞中腺苷酸激活蛋白激酶与核转录因子κBp65的相互作用研究  

Interactions of AMP- activated Protein Kinase( AMPK) and Nuclear Transcription Factor Kappa Bp65 in Adipocytes

作　　者：郑丽英[1] 黄新玲[1] 张焱[1] 李新梅[1] 李静[1] 芦永华[1] 谢建新[1] 

机构地区：[1]石河子大学医学院第一附属医院院内感染控制办,新疆石河子市832000

出　　处：《中国全科医学》2013年第33期3930-3934,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(30960127)

摘　　要：目的探讨腺苷酸激活蛋白激酶(AMPK)与核转录因子κB(NF-κB)p65的相互作用,在细胞水平研究能量代谢与免疫调节间的关系,为阐明肥胖启动炎症反应的分子机制提供实验依据。方法将成纤维细胞3T3-L1诱导为成熟脂肪细胞,取诱导分化第10天的脂肪细胞分为5个组:空白对照组、Compound C组、5氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)组、脂多糖(LPS)组、尼古丁组,分别加入相应试剂。采用蛋白印迹法(Western blotting)检测AMPK与NF-κBp65磷酸化水平,酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子α(TNF-α)与白介素6(IL-6)水平,比色法检测游离脂肪酸(FFA)水平。结果与空白对照组比较,Compound C组AMPK表达水平降低,NF-κBp65表达水平及TNF-α、IL-6、FFA水平升高(P<0.05);AICAR组AMPK表达水平升高,而NF-κBp65表达水平及TNF-α、IL-6、FFA水平降低(P<0.05);LPS组AMPK表达水平降低,而NF-κBp65表达水平升高(P<0.05);尼古丁组AMPK表达水平升高,而NF-κBp65表达水平降低(P<0.05)。结论 AMPK与NF-κBp65之间存在相互作用,两者在能量代谢与免疫调节方面密切相关,肥胖时机体出现的慢性免疫炎症反应可能与AMPK活性降低引发NF-κB信号增强有关。Objective To study the interactions of AMP - activated protein kinase （AMPK） and nuclear transcription factor kappa B （ NF - KB） p65, and the relationship between energy metabolism and immunity adjustment at cell level, in order to provide experimental basis for clarifying the molecular mechanism of obesity - induced inflammation. Methods Fibroblasts 3T3 - L1 was induced to differentiate into mature adipocyte, on the tenth day of differentiation, aidpocytes were divided into the fol- lowing 5 groups： blank control group, Compound C group, AICAR group, LPS group and nicotine group, corresponding rea- gents were added into various groups. Westeron blotting was used to detect the levels of NF - KBp65 and AMPK phosphorylation, ELISA was used to test the expression levels of IL - 6 and TNF - c~, colorimetric method was used to detect FFA level. Results AMPK expression level in Compound C group was significantly lower than that in blank control group, while NF - KBp65 expres- sion level and TNF - α, IL - 6, FFA levels were significantly higher than those in blank control group （ P 〈 0.05 ） . AMPK ex- pression level in AICAR group was significantly higher than that in blank control group, while NF - KBp65 expression level and TNF - α, IL - 6, FFA levels were significantly lower than those in blank control group （ P 〈 0.05 ） AMPK expression level in nicotine group was significantly higher than that in blank control group, while NF - KBp65 expression level was significantly lower than that in blank control group （ P 〈 0. 05 ） . Conclusion There are interactions of AMPK and NF - KBp65, which correlate closely with energy metabolism and immunity adjustment at cell level, chronic immune inflammation in fat body probably corre- lates with NF - KB signal enhancement which is caused by lowered AMPK activity.

关 键 词：脂细胞 腺苷酸激活蛋白激酶 NF-κB 肿瘤坏死因子α 白介素6 脂肪酸类 非酯化 

分 类 号：R349.15[医药卫生—基础医学]