CAJ | 학술논문

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5～40 μmol/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-α EMSA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5～40 μmol/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40 μmol/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-κBB活化.结论 CAI对与炎件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.
목적 탐토최알삼서(CAI)대다충염성상관인자적체외조절작용급궤제.방법 이용MTT법관찰5～40 μmol/L적CAI대세포활력적영향.장상술농도적CAI가입거서세포중공동배양18 h,동시가입지다당(LPS)유도상술세포활화.분별용초산환원법화TNF-α EMSA검측시제합측정배양기상청중NO적함량화TNF-α적수평,용Western blot법측정iNOS단백적표체화NF-κB활화.결과 농도재5～40 μmol/L범위내적CAI대정상대서복강거서세포적세포활력몰유영향,단20화40 μmol/L적CAI능구제량의뢰성지강저LPS유도적NO석방(P＜0.01화P＜0.001)이급TNF-α분비(P＜0.05화P＜0.01),차명현억제LPS인기적iNOS단백표체화NF-κBB활화.결론 CAI대여염건반응상관적조공인자iNOS단백표체화NF-κB활화구유억제작용,해궤제여억제IκBα린산화화강해상관.
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 celia 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 viability 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.