人参皂苷代谢产物Compound K抑制TNF-α诱导的人支气管上皮细胞分泌RANTES  

Ginsenoside metabolite Compound K inhibits TNF-α-induced RANTES secretion in human bronchial epithelial cell line

作　　者：李群益[1,2] 陈莉 张留弟 王轶[1,2] 钟明康[1,2] 施孝金[1,2] 

机构地区：[1]复旦大学附属华山医院临床药学室,上海200040 [2]复旦大学附属华山医院北院临床药学室,上海201907 [3]徐汇区中心医院药械科,上海200031

出　　处：《第二军医大学学报》2015年第7期722-726,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81302853);教育部博士点基金(20110071120071)~~

摘　　要：目的探讨人参皂苷代谢产物Compound K（CK）对炎症因子TNF-α诱导的人支气管上皮细胞BEAS-2B分泌调节活化正常T细胞表达和分泌趋化因子（RANTES）的影响及其机制。方法 ELISA法测定CK对BEAS-2B细胞上清液中RANTES含量的影响;采用RT-PCR和蛋白质免疫印迹技术检测RANTES mRNA及蛋白的表达情况;采用荧光素酶报告基因系统检测CK对活化蛋白转录因子1（activator protein 1,AP-1）和糖皮质激素受体（glucocorticoid receptor,GR）转录抑制或转录激活的影响;运用GR拮抗剂米非司酮,验证CK对RANTES的抑制效应是否由GR介导。结果 3~30μmol/L的CK抑制了TNF-α诱导的RANTES分泌、mRNA及蛋白水平;CK抑制AP-1的转录激活,在BEAS-2B细胞中激活GR信号通路;并且,CK通过GR抑制了BEAS-2B细胞分泌RANTES。结论 CK能够抑制炎症因子TNF-α诱导的支气管上皮细胞分泌RANTES,其机制可能与激活GR、抑制AP-1对下游靶基因的调控有关。Objective To explore the effects of ginsenoside metabolite Compound K（CK）on TNF-α-induced RANTES secretion in human bronchial epithelial cell line BEAS-2Band to elucidate its possible mechanism.Methods BEAS-2Bcells were cultured and treated with CK in different dosages,and then the secretion of RANTES in BEAS-2Bcells exposed to inflammatory stimuli was measured by ELISA kits.Expressions of RANTES mRNA and protein were detected by RT-PCR and Western blotting analysis,respectively.Reporter gene assay was employed to elucidate the interaction between CK and activator protein1（AP-1）,glucocorticoid receptor（GR）.CK antagonist mifepristone was used to observe whether the inhibitory effect of CK against RANTES was mediated by GR.Results TNF-α-induced secretion of RANTES in BEAS-2Bwas markedly inhibited by CK（3-30μmol/L）.Treatment with CK also reduced RANTES mRNA and protein expression.Reporter gene assays indicated that CK was a GR agonist and could repress TNF-α-induced AP-1transactivation.The inhibitory effects of CK on RANTES secretion were antagonized by mifepristone,suggesting apivotal role of GR.Conclusion These results suggest that CK may inhibit TNF-α-induced RANTES secretion in human bronchial epithelial cells,which might be associated with GR pathway activation and AP-1pathway inhibition.

关 键 词：糖皮质激素受体 人参皂苷类 Compound K 炎症 趋化因子CCL5 细胞因子类 哮喘 

分 类 号：R562.25[医药卫生—呼吸系统]