摘要
目的探讨TNF-α和AngⅡ在导致内皮细胞凋亡过程中eNOS的表达变化以及NF-кB在这一过程中的作用.方法对NF-кB的抑制剂PDTC预处理和未预处理的原代培养脐静脉内皮细胞,用TNF-α和AngⅡ分别来进行干预,后用RT-PCR来检测eNOS的mRNA表达,W estern b lot来检测eNOS和IкBα的蛋白表达,EMSA来检测NF-кB的活性,TUNEL法来检测细胞的凋亡.结果在TNF-α和AngII的干预下,eNOS的mRNA和蛋白表达与对照组比较显著下降(P<0.05),细胞凋亡发生显著增加(P<0.05);NF-кB的抑制剂PDTC能抑制TNF-α和AngⅡ引起的细胞凋亡的发生,但未能抑制住TNF-α和AngⅡ引起eNOS表达的下降.结论在TNF-α和AngⅡ作用于内皮细胞时,eNOS和NF-кB对防止细胞凋亡都有保护作用,但NF-кB没有参与TNF-α和AngⅡ引起eNOS表达下降的过程.
Objective To explore the role of eNOS and NF - κB in the cultured endothelial cells apoptosis induced by TNF -α and Ang Ⅱ. Methods The cultured endothelial cells were treated with TNF -α and Ang II in absence and presence of PDTC （ an inhibitor of NF - κB） ; the mRNA of eNOS was measured by RT - PCR, the protein of eNOS and IκBα were assessed by Western - blot, the activity of NF - κB was evaluated by EMSA, and the apoptosis of cells was detected with Tunel. Results The mRNA and proteill of eNOS were significantly decreased in the endothelial cells induced by TNF -α and AngⅡ （ P 〈 0. 05 ） and PDTC couldn t prevent this decrease; TNF -α and Ang Ⅱ could increase the apoptosis of cells （ P 〈 0. 05 ） and PDTC could prevent this increase. Conclusion eNOS and NF -κB both can protect endothelial cells from the apoptosis induced by TNF -α and Ang Ⅱ, but the pathway of NF - κB isnt involved in the decrease of eNOS induced by TNF-α and Ang Ⅱ.
出处
《昆明医学院学报》
2006年第2期13-17,22,共6页
Journal of Kunming Medical College
基金
云南省科委基金资助(2001C0070M)
