异甘草酸镁对成纤维细胞NF-κB信号通路活性的影响  

Effect of magnesium isoglycyrrhizinate on NF-κB signaling pathway activity in fibroblasts

作　　者：徐建国[1] 刘继勇[2] 胡晋红[2] 

机构地区：[1]第二军医大学基础医学部病理生理学教研室,上海200433 [2]第二军医大学长海医院药学部,上海200433

出　　处：《药学服务与研究》2015年第1期22-26,共5页Pharmaceutical Care and Research

基　　金：国家自然科学基金(81373896)

摘　　要：目的:研究异甘草酸镁(magnesium isoglycyrrhizinate,MgIG)对成纤维细胞核因子-kappa B(nuclear factor kappa B,NF-κB)信号通路活性的影响。方法:实验分6组,为空白对照组、TNF-α模型组、地塞米松(dexamethasone,DEX)阳性对照组和0.1、1、10mg/ml的MgIG组。DEX组和MgIG组分别给予DEX(1μg/ml)和MgIG(0.1、1、10mg/ml)预处理4h后,除对照组外,其余5组加入TNF-α(20ng/ml)作用1.5h。提取细胞核蛋白,以免疫印迹法检测NF-κB p65蛋白质表达。提取总RNA,用实时PCR技术检测细胞内IκBαmRNA表达。成纤维细胞经DEX和MgIG预处理4h后加入TNF-α作用24h(浓度同上),用报告基因技术检测NF-κB基因表达。结果:成纤维细胞经TNF-α作用1.5h后,细胞核内NF-κB p65蛋白质表达量明显增加,IκBαmRNA表达上调;TNF-α作用24h后,NF-κB基因上调。DEX预处理4h,可以明显减少TNF-α引起的核内NF-κB p65蛋白质表达和细胞内IκBαmRNA表达,明显降低与TNF-α共作用24h后NF-κB基因表达。MgIG处理成纤维细胞4h后,能够明显降低TNF-α作用1.5h后核内NF-κB p65蛋白质表达量,但对IκBαmRNA表达水平无明显影响。1、10mg/ml的MgIG能够明显降低与TNF-α共作用24h后NF-κB基因表达水平。结论:MgIG的抗炎作用与抑制NF-κB p65转位入核和NF-κB基因表达,从而抑制NF-κB信号通路活性相关,但与DEX不同的是MgIG不能改变IκBαmRNA表达水平。Objective：To investigate the effect of magnesium isoglycyrrhizinate（MgIG）on the activity of nuclear factorkappa B（NF-κB）signaling pathway in fibroblast cells（FCs）.Methods：The experiment consisted of 6groups：the blank control group,the TNF-αmodel group,the dexamethasone（DEX）positive control group,and the 3different concentrations of MgIG groups（i.e.0.1,1and 10mg/ml subgroups）.The DEX group and the MgIG groups were pretreated respectively with DEX（1μg/ml）and MgIG（0.1,1and 10mg/ml）for 4h.Then,with the exception of the control group,all the other 5groups were treated with TNF-α（20ng/ml）for 1.5h.Nuclear proteins and total RNA were collected for detecting the expression levels of NF-κB p65 protein by Western blot.Total RNA was collected and used for detecting the expression levels of IκBαmRNA by real-time PCR.Following pretreatment with DEX and MgIG for 4h,the FCs were treated with TNF-α（with the same dosage as the above）for 24 h,and then the expression levels of NF-κB were detected by reporter gene technology.Results：Following treatment with TNF-αfor 1.5h,the expression levels of intranuclear NF-κB p65 and intracellular IκBαmRNA in FCs were obviously increased,and 24-h treatment with TNF-αalso increased the expression levels of NF-κB.Pretreatment with DEX for 4hcould decrease the expression levels of intranuclear NF-κB p65 and intracellular IκBαmRNA gene induced by TNF-α,and could also significantly reduce the expression levels of NF-κB following combined treatment with TNF-αfor 24 h.Treatment of FCs with MgIG for 4hcould significantly lower the expression levels of intranuclear NF-κB p65 after pretreatment with TNF-αfor 1.5h.However,no significant effects could be noted on the expression levels of IκBαmRNA.MgIG with dosages of 1.0and 10mg/ml could significantly decrease expression levels of NF-κB following combined treatment with TNF-αfor 24 h.Conclusion：The anti-inflammatory effect of MgIG was related with the inhibited NF-κB p65 nuclear trans

关 键 词：异甘草酸镁 NF-ΚB信号通路 抗炎作用 

分 类 号：R285.5[医药卫生—中药学] R966[医药卫生—中医学]