黏着斑激酶抑制剂降低中心静脉导管给药诱导的受损血管内皮细胞与单核细胞黏附的分子机制

Molecular mechanism of focal adhesion kinase inhibitor reducing the adhesion of damaged vascular endothelial cells and monocytes induced by central venous catheter administration

目的研究黏着斑激酶(FAK)抑制剂(Y15)降低中心静脉导管(CVC)给药诱导的血管内皮细胞与单核细胞黏附的分子机制。方法将体外EA.hy926细胞随机分为3组:正常对照组、模型组和实验组。模型组:EA.hy926细胞划痕后与截取好的CVC和THP-1细胞共培养,培养液中加入5-氟脲嘧啶(40μg·mL^(-1));实验组:在模型组基础上加入Y15(50μmol·L^(-1))。蛋白质印迹法测定磷酸化黏着斑激酶Y397(FAKpY397)和锌指转录因子4(GATA4)的表达水平;细胞黏附实验测定THP-1细胞黏附情况。结果干预72 h后,正常对照组、模型组和实验组的FAKpY397的相对表达量分别为0.14±0.03,2.31±0.09和1.27±0.04;这3组的GATA4蛋白相对表达量分别为0.33±0.01,4.75±0.36和1.70±0.08;这3组的THP-1细胞黏附数分别为(2.00±0.71),(41.20±3.03)和(6.00±1.58)个。上述指标:模型组与正常对照组比较,差异均有统计学意义(均P<0.001);实验组与模型组比较,差异均有统计学意义(均P<0.001)。结论黏着斑激酶抑制剂Y15可能通过抑制FAK活化,降低GATA4水平,抑制THP-1细胞的黏附。

Objective To explore the molecular mechanism of focal adhesion kinase(FAK)inhibitor(Y15)in reducing the adhesion of vascular endothelial cells and monocytes induced by central venous catheter(CVC)administration.Methods EA.hy926 cells were randomly divided into 3 groups:normal control group,model group and experimental group in vitro.Model group:EA.HY926 cells were scratched and co-cultured with intercepted CVCs and THP-1 cells,and 5-fluorouracil(40μg·mL^(-1))was added into the culture medium.Experimental group:Y15(50μmol·L^(-1))was added to the model group.The expression levels of phosphorylated focal adhesion kinase Y397(FAK pY397)and zinc finger transcription factor 4(GATA4)were determined by Western blot.The adhesion of THP-1 cells was determined by cell adhesion assay.Results After 72 h of intervention,the relative expression levels of FAK p Y397 in the normal control group,model group and experimental group were 0.14±0.03,2.31±0.09,1.27±0.04,respectively;the relative expression levels of GATA4 protein in the three groups were 0.33±0.01,4.75±0.36,and 1.70±0.08,respectively;the adhesion numbers of THP-1 cells in the three groups were(2.00±0.71),(41.20±3.03)and(6.00±1.58)cell,respectively.Comparison between model group and normal control group,the difference of indicators were significant(all P<0.001);comparison between experimental group and model group except the levels of CORT,the difference of indicators were significant(all P<0.001).Conclusion The focal adhesion kinase inhibitor Y15 may reduce the level of GATA4 and inhibit the adhesion of THP-1 cells by inhibiting FAK activation.