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 10 ng/mL TNF-αfor 0,12,24 and 48 h,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 10 ng/mL TNF-αfor 24 h.The effect of berberine on the prolif-eration of HepG2 cells was assayed by MTT.The level of VEGF mRNA was detected by real-time PCR,and the expression lev-els 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-in-duced VEGF expression at mRNA and protein levels.TNF-αsignificantly decreased the expression of IκBαand increased the ex-pression 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.%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(vascu-lar 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10 ng/mL 的TNF-α分别作用人肝癌细胞(HepG2)0、12、24、48 h；第2部分：分别以0、10、25、50μmol/L 的黄连素预孵育 HepG2细胞24 h后,继续与TNF-α共孵育24 h。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。
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