淫羊藿苷对CX3CL1诱导的单核细胞向人脐静脉内皮细胞迁移和黏附的影响及机制研究

Effects and mechanisms of icariin on migration and adhesion of THP-1 to HUVECs induced by CX3CL1

目的研究淫羊藿苷(icariin,ICA)对CX3CL1诱导的单核细胞(THP-1)向人脐静脉内皮细胞(HUVECs)迁移和黏附的影响及其机制。方法采用Transwells联合培养THP-1和HUVECs,CX3CL1诱导THP-1向HUVECs迁移和黏附模型,并观察ICA(10^(-8)、10^(-7)、10^(-6)mol/L)对该模型的影响。ICA预给药4 h后加入CX3CL1(50 ng/m L)继续培养24 h,移除Transwells及上室中的THP-1细胞,收集培养液并对迁移至下室的THP-1计数;免疫荧光双标法检测THP-1向HUVECs黏附的情况;Western blot检测CX3CR1、p-NF-κB p65和TNF-α蛋白水平。结果与对照组比较,CX3CL1诱导THP-1向HUVECs迁移和黏附的数量明显增多(P<0.01),伴有CX3CR1、p-NF-κB p65、TNF-α蛋白水平升高(P<0.05);ICA预处理(10^(-8)~10^(-6)mol/L)能减少THP-1向HUVECs迁移和黏附的数量(P<0.05或P<0.01),抑制CX3CR1、p-NF-κB p65、TNF-α蛋白水平的上调。结论 ICA(10^(-8)~10^(-6)mol/L)可抑制CX3CL1诱导的THP-1向HUVECs的迁移和黏附,作用机制可能与抑制CX3CL1/CX3CR1/NF-κB/TNF-α信号通路有关。

Objective To investigate the effects and mechanisms of icariin（ ICA） on migration and adhesion of human acute monocytic leukemia cell line（ THP-1） to human umbilical vein endothelial cells（ HUVECs） induced by CX3CL1. Methods Transwells were employed to establish the co-culture system of THP-1 and HUVECs,and to measure the migration and adhesion of THP-1 to HUVECs induced by CX3CL1. After 4 h of ICA administration,CX3CL1（ 50 ng/m L） was added to co-culture system for 24 h. Then the upper chambers of transwells were moved,and the medium was collected for counting the number of the migrated THP-1. The number of adherent THP-1 to HUVECs was observed by immunofluorescence double labeling. Western blot technology was used to evaluate the protein level of CX3CR1,p-NF-κB p65 and TNF-α. Results Compared with the control group,the number of migration and adhesion of THP-1 to HUVECs induced by CX3CL1 was significantly increased（ P 〈 0. 01）,and the protein levels of CX3CR1,p-NF-κB p65 and TNF-α were also increased（ P 〈 0. 05）. Pretreatment with ICA could cut down the number of migration and adhesion of THP-1 to HUVECs in dose-dependent manner at the dosage range from 10-8mol/L to 10-6mol/L（ P 〈 0. 05 or P 〈 0. 01）,and inhibit the protein levels of CX3CR1,p-NF-κB p65 and TNF-α. Conclusion ICA（ 10-8 10-6mol/L） inhibit the migration and adhesion of THP-1 to HUVECs induced by CX3CL1 through inhibition of CX3CL1/CX3CR1/NF-κB/TNF-α signaling pathway.