CX3CL1介导人脐静脉内皮细胞骨架改变的功能研究

发布时间：2018-07-21 11:11

【摘要】：该研究探讨了趋化因子CX3CL1对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)骨架的影响及其作用机制。CX3CL1刺激HUVECs后,采用免疫荧光染色技术检测细胞骨架蛋白纤维状肌动蛋白(F-actin)的分布和形态改变,采用Western blot技术检测胞质内F-actin和磷酸化促分裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)的三种亚型[p38、细胞外调节蛋白激酶1/2(extracellular regulated protein kinase 1/2,ERK1/2)和c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)]的表达水平。结果显示,10 nmol/L CX3CL1刺激HUVECs 30 min后,细胞的致密外周带逐渐被破坏,胞质内有应力纤维形成;120 min后,外周带消失,胞质内有大量的致密应力纤维形成;180 min后,胞质内应力纤维减少,少数细胞可见致密外周带。10 nmol/L CX3CL1刺激HUVECs30 min后,F-actin的表达水平逐渐升高,并于120 min后达峰值;10 nmol/L CX3CL1刺激HUVECs 1 min后,磷酸化p38、ERK1/2和JNK表达水平升高,5 min后三者的表达水平达峰值;5μg/m L抗CX3CR1抗体抑制10 nmol/L CX3CL1刺激HUVECs后,磷酸化p38、ERK1/2和JNK的表达水平降低;30μmol/L p38的特异性抑制剂SB203580和ERK1/2的特异性抑制剂PD98059抑制10 nmol/L CX3CL1刺激HUVECs后,胞质内应力纤维减少,应力纤维变短,F-actin的表达水平降低。以上研究结果表明,CX3CL1能通过p38和ERK1/2信号通路以时间依赖方式介导HUVECs细胞骨架的重构。
[Abstract]:The aim of this study was to investigate the effect of CX3CL1 on the cytoskeleton of human umbilical vein endothelial cells (human umbilical vein endothelial cells) and its mechanism. The distribution and morphology of F-actin, a cytoskeleton protein, were detected by immunofluorescence staining after stimulation of HUVECs by CX3CL1. Three subtypes of intracellular F-actin and phosphorylated mitogen-activated protein kinase (mitogen activated protein kinaseshs) [p38, extracellular regulated protein kinase 1 / 2 (extracellular regulated protein kinase 1 / 2 ERK1 / 2) and c-Jun N-terminal kinase (JNK) were detected by Western blot assay. The results showed that after 10 nmol / L CX3CL1 stimulated HUVECs for 30 min, the dense peripheral zone of the cells was destroyed gradually. After the stress fibers were formed in the cytoplasm for 120 min, the peripheral bands disappeared, and a large number of dense stress fibers formed in the cytoplasm for 180 min, and the endoplasmic stress fibers decreased. In a few cells, the expression of F-actin in HUVECs was gradually increased after stimulation of HUVECs 30 min with dense peripheral zone of .10 nmol / L CX3CL1, and reached a peak value of 10 nmol / L CX3CL1 at 120 min after stimulation of HUVECs 1 min. The expression of phosphorylated p38 ERK1 / 2 and JNK in HUVECs increased to a peak of 5 渭 g / mL after 5 min. The anti-CX3CR1 antibody inhibited the stimulation of HUVECs by 10 nmol / L CX3CL1. The expression level of phosphorylated p38 ERK1 / 2 and JNK decreased by 30 渭 mol / L p38, a specific inhibitor SB203580 and ERK1 / 2, and PD98059 inhibited the expression of stress fibers and F-actin in HUVECs stimulated by 10 nmol / L CX3CL1. These results suggest that CX3CL1 can mediate the cytoskeleton remodeling of HUVECs in a time-dependent manner through p38 and ERK1 / 2 signaling pathways.
【作者单位】：重庆医科大学附属第二医院肾内科;
【基金】：重庆市卫生局医学科研项目(批准号:2012-1-034)资助的课题~~
【分类号】：R329.2

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