青藤碱抑制肿瘤坏死因子α刺激引起的人脐静脉内皮细胞核因子κB p65的核移位
Sinomenine inhibited TNF-α induced NF-κB p65 level in human umbilical vein endothelial cells nucleus
目的 研究青藤碱(SIN)对肿瘤坏死因子α(TNF-α)刺激引起的人脐静脉内皮细胞(HUVEC)核因子(NF)κB p65的核移位的影响.方法 从新鲜脐带中分离并培养HUVEC,用TNF-α诱导NF-κB p65核移位及血管细胞黏附分子1(VCAM-1)表达.实验组加入不同浓度的SIN(0.25、05和1.0 mol/L)或地塞米松(Dex,1.0×10-6 mol/L)进行干预,收获细胞.提取细胞总核蛋白,用ELISA法检测核提取物中NF-κB p65水平;用实时定量PCR检测VCAM-1 mRNA 的表达.结果 所提取的核蛋白浓度在0.16 g/L至0.77 g/L之间.与TNF-α刺激组相比,SIN干预组(0.25 mol/L、050 mol/L 和1.0 mol/L)和Dex (1.0×10-6 mol/L)干预组核提取物中NF-κB p65水平下降,各干预组VCAM-1 mRNA相对表达量有不同程度下降(P<0.05).结论 不同浓度的SIN(0.25、0.5、1.0 mol/L)和Dex (1.0×10-6 mol/L)能抑制TNF-α诱导的人脐静脉内皮细胞的NF-κB p65核转移.
更多Objective To investigate the effect of sinomenine(SIN)on TNF-α induced NF-κB p65 level in human umbilical vein endothelial cells(HUVECs)nucleus.Methods HUVECs were isolated from freshly collected umbilical cords. Positive controls samples were stimulated with TNF-α,omitting SIN. Negative controls samples were treated in the same way,omitting TNF-α and SIN. Experiment samples were co-cultured with TNF-α and SIN at different concentrations (0.25,0.5,and 1.0 mol/L),or TNF-α and dexamethasone(Dex) at concentration of 1.0×10-6 mol/L.Cell nuclear extracts were prepared according to extraction procedure of Nuclear Extraction Kit .NF-κB p65 in cells nuclear extracts was detected by ELISA.Results The protein concentration of cell nuclear extracts samples were from 0.16 g/L to 0.77 g/L. NF-κB p65 levels in cell nucleus in the groups of SIN (at concentration of 0.25 mol/L to 1.0 mol/L) and Dex (at concentration of 1.0×10-6 mol/L) decreased compared with the group of positive control.Conclusion SIN and Dex inhibit NF-κB p65 level of cell nucleus in HUVECs stimulated by TNF-α in vitro.
More- 浏览:143
- 被引:0
- 下载:8
相似文献
- 中文期刊
- 外文期刊
- 学位论文
- 会议论文