高糖诱导人内皮细胞凋亡及其JNK、AKT信号途径的作用机制  

High level of glucose induced apoptosis in human endothelial cells and its mechanism of JNK and AKT signaling pathways

作　　者：刘云峰[1] 杨川[1] 程桦[1] 黎明涛[2] 

机构地区：[1]中山大学第二医院内分泌科,广东广州510120 [2]中山大学基础医学院药理教研室,广东广州510080

出　　处：《基础医学与临床》2005年第2期163-168,共6页Basic and Clinical Medicine

摘　　要：目的探讨高糖诱导人脐静脉内皮细胞(humanumbilicalveinendothelialcells,HUVECs)凋亡的JNK及AKT信号途径作用机制。方法HUVECs细胞分别在生理浓度葡萄糖(5mmol/L) (NG)、高葡萄糖(30mmol/L) (HG)、高糖加JNK特异性阻断剂SP6 00 12 5 (10 μmol/L) (HG +I)条件下培养72h。Hoechst332 5 8染色,荧光显微镜观察细胞凋亡的形态学改变;annexinV FITC试剂盒染色,流式细胞仪进行细胞凋亡定量;Westernblot方法测定细胞p JNK、p- c- JUN、p AKT水平。结果高葡萄糖培养72h ,内皮细胞凋亡率为13 .31% ,显著高于对照组5 . 6 9% (P <0 . 0 1)。高糖条件下内皮细胞p JNK、p c JUN水平较对照组显著增加(P <0 . 0 5 ) ,而p AKT显著降低(P <0 . 0 1)。SP6 0 0 12 5应用后,与高糖组比较p JNK无显著变化、p c JUN含量降低(P <0 .0 5 ) ,而p AKT升高(P <0 . 0 1) ,高糖诱导的内皮细胞凋亡被抑制(8. 38% ,P <0 . 0 1)。结论高糖可诱导内皮细胞凋亡,其机制可能是激活JNK及其下游c- JUN ,而抑制AKT的活化,而JNK活化程度对AKT信号通路可能存在调节作用。Objective To investigate the signaling pathway of high level of glucose-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were incubated with physiological concentration of glucose (5 mmol/L) (NG), high glucose (30 mmol/L)(HG) and high glucose combined with SP600125(HG+I) for 72 hours. Apoptosis of HUVECs was observed in a fluorescence microscope by Hoechst 33258 stain, and quantitied by flow cytometry using annexin V-FITC stain. The level of p-JNK, p-c-JUN and p-AKT was evaluated by western blot. Results Treatment of HUVECs with high glucose resulted in a significant increase in apoptotic cells comparing to a physiological concentration (13.31% vs 5.69%, P<0.01). Incubation of HUVECs with high concentration of glucose for 72 hours increased the phosphorylation of JNK and c-JUN significantly (P<0.05, vs NG), while decreased the level of p-AKT markedly(P<0.01, vs NG). After treated with SP600125, the level of p-JNK and p-c-JUN (P<0.05, vs HG) decreased, while the level of p-AKT increased (P<0.01, vs HG),and the apoptosis of HUVECs induced by high glucose was inhibited (8.38%, P<0.01, vs HG). Conclusion High concentration of glucose could induce HUVECs apoptosis, probably through increasing the activation of JNK, the downstream c-jun, and inhibition of the activation of AKT. The activation JNK could regulate AKT pathway.

关 键 词：JNK 高糖 C-JUN 诱导 内皮细胞凋亡 HUVEC 人内皮细胞 信号途径 N含量 阻断剂 

分 类 号：R587.1[医药卫生—内分泌]