刀豆蛋白A诱导人脐静脉内皮细胞表达p-ERK1/2

Concanavalin A enhances p-ERK1/2 expression in CRL2480 cells

目的：探讨刀豆蛋白A （Concanavalin A，ConA）对CRL2480细胞（人脐静脉内皮细胞系）细胞外调节蛋白激酶（Extracellular signal-regulated kinase，ERK）1/2磷酸化的影响。方法采用MTT法检测ConA对CRL2480细胞活化的影响；采用Western印迹法检测CRL2480细胞磷酸化（p）-ERK1/2的表达；采用细胞免疫荧光技术检测p-ERK1/2在细胞内的分布。结果在5~20μg/ml范围，ConA刺激CRL2480细胞8 h的活化指数具有剂量依赖性；20μg/ml ConA显著增加CRL2480细胞的活化（P〈0.05）。ConA能够增加CRL2480细胞的p-ERK1/2表达水平（P〈0.05），ERK途径抑制剂PD98059和U0126明显抑制p-ERK1/2表达。免疫荧光染色结果显示ConA刺激细胞的荧光信号明显高于对照细胞，荧光分布于细胞浆、细胞核周围和细胞核内。结论 ConA能够激活CRL2480细胞表达p-ERK1/2。

Objective To explore the influence of concanavalin A （ConA） on the expression of phosphorylated extra-cellular signal-regulated kinase （p-ERK） 1/2 in CRL2480 cells （human umbilical vein endothelial cell line）. Methods Activat-ed effect of ConA on CRL2480 endothelial cells was detected by MTT assay. P-ERK1/2 expression was examined by western blot. The distribution of p-ERK1/2 was observed via immunofluorescent staining. Results After 8h incubation, ConA in-creased the activated index of the cells in a dose dependent manner in the range of 5~20μg/ml （P〈0.05）. The immu-noblot displayed that 20 μg/ml ConA increased p-ERK1/2 expression in the cells （P〈0.05）. PD98059 and U0126 （both ERK pathway inhibitors） exerted inhibitory action on p-ERK1/2 expression （P〈0.05）. The fluo-rescent intensities existed in the cytoplasm and the nuclei of ConA-treated cells were much stronger than those of control cells. Conclusion ConA is able to upregulate p-ERK1/2 expression in CRL2480 cells.