羧胺三唑抑制大鼠腹腔巨噬细胞iNOS表达和NF-κB活化  

Inhibitory effect of CAI on iNOS expression and NF-κB activation degradation in rat peritoneal macrophages

作　　者：郑茹[1,2] 郝晓健[1,2] 郭磊[1,2] 李娟[1,2] 于晓丽[1,2] 叶菜英[1,2] 张德昌[1,2] 

机构地区：[1]中国医学科学院基础医学研究所 [2]北京协和医学院基础医学院药理室,北京100005

出　　处：《基础医学与临床》2010年第5期466-470,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30873075);国家科技重大专项(2009ZX09303-008);"重大新药创制"科技重大专项(2009ZX09102-049);院所长基金(2009PY01)

摘　　要：目的探讨羧胺三唑（CAI）对多种炎性相关因子的体外调节作用及机制。方法利用MTT法观察5-40μmol/L的CAI对细胞活力的影响。将上述浓度的CAI加入巨噬细胞中共同培养18 h,同时加入脂多糖（LPS）诱导上述细胞活化。分别用硝酸还原法和TNF-αELISA检测试剂盒测定培养基上清中NO的含量和TNF-α的水平,用Western blot法测定iNOS蛋白的表达和NF-κB活化。结果浓度在5-40μmol/L范围内的CAI对正常大鼠腹腔巨噬细胞的细胞活力没有影响,但20和40μmol/L的CAI能够剂量依赖性地降低LPS诱导的NO释放（P〈0.01和P〈0.001）以及TNF-α分泌（P〈0.05和P〈0.01）,且明显抑制LPS引起的iNOS蛋白表达和NF-κB活化。结论CAI对与炎性反应相关的调控因子iNOS蛋白表达和NF-κB活化具有抑制作用,该机制与抑制IκBα磷酸化和降解相关。Objective This study is designed to explore the regulation of carboxyamidotriazole（CAI） on inflammatory factors in vitro and its underlying mechanisms.Methods Peritoneal macrophages were incubated with different concentrations（5～40 μmol/L）,and then cell viabilities were evaluated by MTT assay.Cells were pretreated with CAI（5～40 μmol/L）,LPS was then added,and cells were incubated for 18 h.NO and TNF-α levels were determined with Griess reagent and ELISA kit,respectively.The iNOS expression and NF-κB activation were detected by Western blot method.Results The rat peritoneal macrophage viablilty was not affected at the concentrations of CAI used.CAI（5～40 μmol/L） was found to reduce NO（P〈0.01,P〈0.001） and TNF-α production（P〈0.05,P〈0.01） in a dose-dependent manner.CAI was also found to inhibit the LPS-induced expression of iNOS and degradation of IκBα in rat peritoneal macrophages.Conclusion The findings from the present study suggest that CAI has suppressive effect on iNOS expression,and this inhibitory effect was found to be associated with NF-κB inactivation via the blockade of IκBα phosphorylation.

关 键 词：羧胺三唑(CAI) 巨噬细胞 INOS NF-ΚB活化 

分 类 号：R285.5[医药卫生—中药学]