海南医学
海南醫學
해남의학
HAINAN MEDICAL JOURNAL
2014年
14期
2029-2031,2032
,共4页
龚文广%王红梅%牛青霞
龔文廣%王紅梅%牛青霞
공문엄%왕홍매%우청하
刀豆球蛋白A%内皮细胞%细胞外调节蛋白激酶
刀豆毬蛋白A%內皮細胞%細胞外調節蛋白激酶
도두구단백A%내피세포%세포외조절단백격매
Concanavalin A%Endothelial Cells%Extracellular signal-regulated kinase
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。
目的:探討刀豆蛋白A (Concanavalin A,ConA)對CRL2480細胞(人臍靜脈內皮細胞繫)細胞外調節蛋白激酶(Extracellular signal-regulated kinase,ERK)1/2燐痠化的影響。方法採用MTT法檢測ConA對CRL2480細胞活化的影響;採用Western印跡法檢測CRL2480細胞燐痠化(p)-ERK1/2的錶達;採用細胞免疫熒光技術檢測p-ERK1/2在細胞內的分佈。結果在5~20μg/ml範圍,ConA刺激CRL2480細胞8 h的活化指數具有劑量依賴性;20μg/ml ConA顯著增加CRL2480細胞的活化(P<0.05)。ConA能夠增加CRL2480細胞的p-ERK1/2錶達水平(P<0.05),ERK途徑抑製劑PD98059和U0126明顯抑製p-ERK1/2錶達。免疫熒光染色結果顯示ConA刺激細胞的熒光信號明顯高于對照細胞,熒光分佈于細胞漿、細胞覈週圍和細胞覈內。結論 ConA能夠激活CRL2480細胞錶達p-ERK1/2。
목적:탐토도두단백A (Concanavalin A,ConA)대CRL2480세포(인제정맥내피세포계)세포외조절단백격매(Extracellular signal-regulated kinase,ERK)1/2린산화적영향。방법채용MTT법검측ConA대CRL2480세포활화적영향;채용Western인적법검측CRL2480세포린산화(p)-ERK1/2적표체;채용세포면역형광기술검측p-ERK1/2재세포내적분포。결과재5~20μg/ml범위,ConA자격CRL2480세포8 h적활화지수구유제량의뢰성;20μg/ml ConA현저증가CRL2480세포적활화(P<0.05)。ConA능구증가CRL2480세포적p-ERK1/2표체수평(P<0.05),ERK도경억제제PD98059화U0126명현억제p-ERK1/2표체。면역형광염색결과현시ConA자격세포적형광신호명현고우대조세포,형광분포우세포장、세포핵주위화세포핵내。결론 ConA능구격활CRL2480세포표체p-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.