IL-18激活NF-κB细胞信号通路对人脐静脉内皮细胞损伤作用的体外研究

发布时间：2018-08-28 19:27

【摘要】：目的探讨促炎因子白细胞介素-18(IL-18)通过激活核因子-κB(NF-κB)介导的细胞信号通路,对静脉内皮细胞功能的影响及其与深静脉血栓形成(DVT)的联系。方法利用重组人IL-18作用体外培养的人脐静脉内皮细胞(HUVECs),并以NF-κB激活抑制剂进行干预,通过实时荧光定量PCR、Western blot、免疫荧光、流式细胞仪等检测手段,验证IL-18是否通过激活NF-κB介导的细胞信号转导通路,影响HUVECs正常状态及血管性血友病因子(vWF)、P-选择素(P-selectin)、组织型纤溶酶原激活物(t-PA)等内皮细胞功能标记物的表达,结合既往研究对IL-18参与DVT的机制进行综合分析。结果 IL-18可激活内皮细胞内NF-κB,使细胞核内p65表达增高、细胞内IκBα表达降低,并使HUVECs早期凋亡细胞明显增多;添加QNZ(EVP4593)可使IL-18对NF-κB的激活作用明显抑制,细胞损伤、凋亡的发生显著减少;IL-18可促使vWF、P-selectin和t-PA等DVT相关的内皮细胞标志物发生表达异常(P0.05),而各标志物可在NF-κB激活抑制后恢复常规表达。结论 IL-18及NF-κB间的相互作用导致HUVECs生长状态和功能异常,可能是与DVT发病相关的疾病机制。
[Abstract]:Objective to investigate the effect of interleukin-18 (IL-18) on the function of vascular endothelial cells (VECs) by activating nuclear factor- 魏 B (NF- 魏 B) -mediated cellular signaling pathway and its association with deep venous thrombosis (DVT) (DVT). Methods Human umbilical vein endothelial cells (HUVECs),) cultured in vitro were treated with recombinant human IL-18 and interfered with NF- 魏 B activator. Real-time fluorescence quantitative PCR,Western blot, immunofluorescence and flow cytometry were used to detect (HUVECs),. To verify whether IL-18 affects the expression of vascular Willebrand factor (vWF) P- selectin (P-selectin), tissue plasminogen activator (t-PA) and other endothelial cell functional markers by activating NF- 魏 B mediated cell signal transduction pathway. Combined with previous studies, the mechanism of IL-18 participating in DVT was comprehensively analyzed. Results IL-18 could activate NF- 魏 B in endothelial cells, increase the expression of p65, decrease the expression of I 魏 B 伪 and increase the number of early apoptotic cells in HUVECs, and the addition of QNZ (EVP4593) could inhibit the activation of NF- 魏 B and damage the cells. The decrease of apoptosis could promote the abnormal expression of DVT related endothelial cell markers such as vWF,P-selectin and t-PA (P0.05), and the markers could return to normal expression after NF- 魏 B activation inhibition. Conclusion the interaction between IL-18 and NF- 魏 B leads to abnormal growth and function of HUVECs, which may be related to the pathogenesis of DVT.
【作者单位】：贵州医科大学附属医院骨科;昆明医科大学第一附属医院骨科;
【基金】：国家自然科学基金面上项目(30960389) 云南省卫生科技计划项目(2014NS141) 云南省重点新产品开发计划项目(2010BC010) 云南省卫计委内设机构项目(2016NS021) 贵州医科大学博士基金(院博合J字[2015]019号) 贵州省科技厅合作计划项目(黔科合LH字[2016]7228号)
【分类号】：R543.6

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