黄连素通过NF-κB信号通路对TNF-α诱导的VEGF表达的影响  

Effect of Berberine on TNF-α-induced VEGF Expression via NF-κB Signaling Pathway

作　　者：李井彬[1] 柯善栋[2] 胡少明[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院中西医结合科,武汉430030 [2]湖北省黄石市中心医院血液科,黄石435000

出　　处：《华中科技大学学报（医学版）》2014年第4期386-390,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：科技部国际科技合作项目(No.2010DFA31870)

摘　　要：目的观察黄连素对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导的血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达的影响及探讨其可能机制。方法实验分两部分,第1部分:以10ng/mL的TNF-α分别作用人肝癌细胞(HepG2)0、12、24、48h;第2部分:分别以0、10、25、50μmol/L的黄连素预孵育HepG2细胞24h后,继续与TNF-α共孵育24h。MTT法检测黄连素对HepG2细胞增殖能力的影响;Real-time PCR检测各组细胞VEGF mRNA水平;Western blot检测各组细胞VEGF、IκBα、NF-κB p65蛋白表达;ELISA检测NF-κB的活性。结果黄连素对HepG2细胞生长的抑制能力呈浓度依赖性;TNF-α显著上调HepG2细胞中VEGF mRNA和蛋白表达且呈时间依赖性;黄连素显著下调TNF-α诱导的HepG2细胞中VEGF mRNA和蛋白表达且呈浓度依赖性;TNF-α显著下调IκBα表达且呈时间依赖性,上调NF-κB P65的表达及NF-κB活性且呈时间依赖性;黄连素显著上调IκBα表达且呈浓度依赖性,下调NF-κB P65的表达及NF-κB活性且呈浓度依赖性。结论 TNF-α在HepG2细胞中能上调VEGF的表达,而黄连素能下调由其诱导的VEGF表达,可能是通过NF-κB信号通路所介导的。Objective To examine the effect of berberine on tumor necrosis factor-α（TNF-α）-induced VEGF expression and the possible mechanism.Methods There were two sessions in this study.In the first session,HepG2 cells were treated with 10ng/mL TNF-αfor 0,12,24 and 48h,respectively.In the second session,HepG2 cells were pre-incubated with 0,10,25 and 50μmol/L berberine for 24 h,respectively,and then treated with 10ng/mL TNF-αfor 24 h.The effect of berberine on the proliferation of HepG2 cells was assayed by MTT.The level of VEGF mRNA was detected by real-time PCR,and the expression levels of VEGF,IκBαand NF-κB p65 proteins by Western blot.The activity of NF-κB was measured by ELISA.Results Berberine could dose-dependently inhibit the growth of HepG2 cells.TNF-αcould increase the mRNA and protein expression levels of VEGF in HepG2 cells in a time-dependent manner.Berberine could significantly dose-dependently down-regulate the TNF-α-induced VEGF expression at mRNA and protein levels.TNF-αsignificantly decreased the expression of IκBαand increased the expression of NF-κB p65 and the activity of NF-κB in a time-dependent manner,which could be dose-dependently abrogated by pretreatment with berberine.Conclusion Berberine inhibits TNF-α-induced VEGF expression via NF-κB signaling pathway.

关 键 词：黄连素 HEPG2 血管内皮细胞生长因子 肿瘤坏死因子-α 核转录因子-ΚB 

分 类 号：R282.71[医药卫生—中药学]