核因子-κB在HUVEC细胞凋亡信号通路中的作用

目的：探讨核因子-κB（NF-κB）在人脐静脉内皮细胞凋亡信号通路中的作用。方法：体外培养人脐静脉内皮细胞系（HUVEC）,实验分为正常对照组、AngⅡ组和Gliotoxin干预组。应用改良MTF法,观察0．01μmol／L、0．1μmol／L、μmol／L和10μmol／L4种浓度的AngⅡ在不同时间对HUVEC细胞活性的影响。应用DNA凝胶电泳和流式细胞术检测AngⅡ作用于细胞后引起细胞凋亡的情况。应用免疫细胞化学技术检测NF-κB p65的核移位,评价NF-KB活化情况。结果：10μmol／L AngⅡ作用于细胞24h时,细胞活性下降,DNA凝胶电泳和流式细胞结果提示细胞发生凋亡,凋亡细胞率明显高于正常对照组,差异具有统计学意义（P〈0．05）,0．1mg／L Gliotoxin可拮抗AngⅡ的细胞抑制活性作用;免疫细胞化学技术显示,HUVEC细胞经AugⅡ诱导后,NF-κB出现明显核移位现象,提示NF-κB发生活化;Gliotoxin明显抑制NF-κB活化,与AngⅡ组相比,差异有统计学意义（P〈0．05）。结论：①ArcⅡ可引起HU—VEC细胞发生凋亡;而NF-κB特异性抑制剂Ghotoxin能够拮抗AngⅡ对HUVEC细胞的作用;②NF-κB可能是AngⅡ调控HUVEC细胞生存／凋亡通路中的重要信号转导分子。

Role of nuclear factor-kappaB in apoptosis pathway of HUVEC

Aim： To investigate the role of nuclear factor-κB in apoptosis pathway of HUVEC. Methods： The cell lines of HUVEC cultured in vitro were divided into three groups： normal control group, Ang Ⅱ group, and Gliotoxin group. We investigated the effects of Ang Ⅱ （0.01μmol/ L, 0. 1 μmol/L, 1 μmol/L and 10 μmol/L） on the viability of HUVEC with modified MTT.Then agarose gel electrophoresis and flow cytometry were applied to detect the apoptosis of HUVEC. Finally, the nuclear translocation of NF-κB subunit p65 was evaluated by immunocytochemistry. Results： The viability of HUVEC decreased significantly after incubated with 10 μmol/L Ang Ⅱ for 24 hours. The results of DNA agarese gel and flow cytometry showed that 10 μmol/L Ang Ⅱ induced the apoptosis of HUVEC, and the apoptosis rate was significantly higher than normal control group（ P 〈 0.05）. 0.1 mg/L Gliotoxin antagonized this effect of Ang Ⅱ. The results of immunocytochemistry suggested that NT-κB was activated in HUVEC induced by 10 μmol/L Ang Ⅱ. In contrast, Gliotoxin inhibited the activation of NF-κB in HUVEC induced by Ang Ⅱ. Conclusion： ①Ang Ⅱ can induce the apoptosis of HUVEC, while the inhibitor of NF-κB, Gliotoxin, can antagonize the effect of Ang ft. ②NF-κB may play an important role in apoptosis pathway of HUVEC induced by Ang Ⅱ.