Zhao Yongbo, Feng Guangxing, Wang Yanzhi, Yue Yuehong, Zhao Weichao
Department of Cardiovascular Surgery, Fourth Hospital of Hebei Medical University Shijiazhuang, China.
Department of Neurology, Hebei General Hospital Shijiazhuang, China.
Int J Clin Exp Pathol. 2014 Oct 15;7(11):7699-707. eCollection 2014.
This study was performed to investigate PTX3-mediated iNOS expression and IKK/IκB/NF-κB activation in PA-induced atherosclerotic HUVECs injury model.
The cell viability was detected by the CCK8 assay. The cell apoptosis was assessed by annexin V-PI double-labeling staining. Expression of genes and proteins were analyzed by real-time PCR and western blotting respectively. Cells were transfected with siRNAs as a gene silencing methods.
PA induced cell apoptosis in human umbilical vein endothelial cells in a time and dose-dependent manner. PA also induced upregulation expression of PTX3. TPCA-1, an inhibitor of IKK-2, could suppress the expression of PTX3 and phospho-IκB-α in PA-induced endothelial dysfunction cell model. We also found that transfection of cells with PTX3 siRNA reduced the expression of iNOS and NO, and protected PA-induced cell apoptosis in HUVECs.
PTX3 could exacerbate endothelial dysfunction, at least partially, through IKK/IκB/NF-κB activation and overexpression of iNOS and NO, and advance the development of atherosclerosis.
本研究旨在探讨在棕榈酸（PA）诱导的动脉粥样硬化人脐静脉内皮细胞（HUVECs）损伤模型中，长五聚蛋白3（PTX3）介导的诱导型一氧化氮合酶（iNOS）表达及IκB激酶（IKK）/IκB/核因子κB（NF-κB）激活情况。
采用细胞计数试剂盒-8（CCK8）法检测细胞活力。通过膜联蛋白V-碘化丙啶（annexin V-PI）双染法评估细胞凋亡。分别用实时定量聚合酶链反应（real-time PCR）和蛋白质免疫印迹法分析基因和蛋白质表达。以小干扰RNA（siRNAs）转染细胞作为基因沉默方法。
PA以时间和剂量依赖性方式诱导人脐静脉内皮细胞凋亡。PA还诱导PTX3表达上调。IKK-2抑制剂TPCA-1可抑制PA诱导的内皮功能障碍细胞模型中PTX3和磷酸化IκB-α的表达。我们还发现，用PTX3 siRNA转染细胞可降低iNOS表达及一氧化氮（NO）水平，并保护HUVECs免受PA诱导的细胞凋亡。
PTX3可至少部分通过激活IKK/IκB/NF-κB以及iNOS和NO的过表达加重内皮功能障碍，促进动脉粥样硬化发展。
