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目的探讨p38MAPK信号通路特异性抑制剂SB203580对血流低剪切应力诱导的人脐静脉内皮细胞株Fractalkine蛋白表达的影响。方法用M199生长液(含1%的青链霉素合剂和10%的胎牛血清)培养EA.hy926细胞至第2~5代,待细胞长满汇合后加载强度为4.58dyne/cm2的剪切应力,分别处理0、5、10、30、60min。收集细胞提总蛋白,通过Western-blot技术检测剪切应力处理不同时间对EA.hy926细胞p38 MAPK磷酸化的影响,并分析p38 MAPK特异性抑制剂SB203580对p38磷酸化水平的抑制程度,观察SB203580对低剪切应力诱导的人脐静脉内皮细胞株Fractalkine蛋白表达的影响。结果低剪切应力处理可引起EA.hy926细胞p38蛋白磷酸化水平的上调,此过程与剪切应力作用时间有关,30min时磷酸化水平最高。用SB203580处理可明显抑制低剪切应力诱导的EA.hy926细胞Fractalkine蛋白表达上调过程。结论低剪切应力可激活p38信号转导途径,激活的p38信号分子可能参与调节低剪切应力诱导的内皮细胞Fractalkine表达上调过程,p38抑制剂能够下调低剪切应力诱导的Fractalkine的表达。
Objective To investigate the effect of SB203580, a specific inhibitor of p38 MAPK signaling pathway, on the expression of Fractalkine in human umbilical vein endothelial cells induced by low shear stress. Methods EA.hy926 cells were cultured in M199 medium (containing 1% streptomycin and 10% fetal bovine serum) until the second to fifth passages. After the cells were confluent, the cells were loaded with shear strength of 4.58 dyne / cm2 Cut stress, respectively, 0,5,10,30,60 min. The total protein was collected and the effect of shear stress treatment on the phosphorylation of p38 MAPK in EA.hy926 cells was detected by Western-blot. The inhibition of p38 phosphorylation by SB203580, a specific inhibitor of p38 MAPK, was analyzed. The inhibitory effect of SB203580 Effects of Low Shear Stress on Fractalkine Protein Expression in Human Umbilical Vein Endothelial Cell. Results Low shear stress treatment induced the phosphorylation of p38 in EA.hy926 cells. The process was related to the time of shear stress and the highest level of phosphorylation at 30 min. Treatment with SB203580 significantly inhibited the up-regulation of Fractalkine protein expression in EA.hy926 cells induced by low shear stress. Conclusion Low shear stress can activate the p38 signal transduction pathway. The activated p38 signaling molecule may be involved in the regulation of Fractalkine expression in endothelial cells induced by low shear stress. P38 inhibitor can down-regulate the expression of Fractalkine induced by low shear stress.