艾溴利平对高糖诱导人脐静脉内皮细胞凋亡及蛋白激酶B表达的影响  

Effect of ibrolipim on human umbilical vein endothelial cells apoptosis and protein kinase B expression induced by high glucose

作　　者：肖国华[1] 王宗保[2] 余坚[2] 高天舒[2] 梅红霞[2] 贺震[2] 

机构地区：[1]南华大学药学与生命科学学院,湖南衡阳421001 [2]南华大学实验动物部,湖南衡阳421001

出　　处：《中国新药与临床杂志》2010年第8期611-616,共6页Chinese Journal of New Drugs and Clinical Remedies

基　　金：湖南省科技厅科研项目(2008SKJ16)

摘　　要：目的观察艾溴利平对高糖诱导人脐静脉内皮细胞凋亡和蛋白激酶B(PKB)磷酸化的影响,并探讨其分子机制。方法应用终浓度为33.3 mmol·L-1葡萄糖诱导内皮细胞HUVEC-12细胞株高糖模型,实验分为正常对照组、高糖模型组、艾溴利平组和抑制剂组,Hoechst33258染色及AnnexinV和PI双染流式细胞术检测细胞凋亡;Western blotting检测PKB总蛋白、PKB苏氨酸308(PKB-Thr308)及PKB丝氨酸473(PKB-Ser473)的磷酸化水平。结果高糖模型组内皮细胞凋亡指数(AI)增加为(14.07±s 0.25)%,高于正常对照组(P<0.01),艾溴利平8μmo·lL-1组AI降至(5.42±0.13)%,与高糖模型组相比有非常显著差异(P<0.01),抑制剂组AI增加至(12.00±0.10)%。低浓度艾溴利平(2、4、8μmo·lL-1)短时间(6、12、24、48 h)增加PKB磷酸化(P<0.01);而阻断PKB活性可显著抑制艾溴利平的抗凋亡作用。结论艾溴利平能抑制高糖引起的人脐静脉内皮细胞凋亡,可能与信号分子PKB的激活有关。AIM To observe the effect of ibrolipim on the apoptosis and protein kinase B （PKB） phasphorylation in cultured human umbilical vein endothelial cells （HUVECs） induced by high glucose, and to investigate its molecular mechanism. METHODS HUVEC-12 cell strain high glucose model groups were induced by 33.3 mmol·L^-1 D-glucose, and experiments were randomized into four groups： control group, high glucose model group, ibrolipim group, and inhibitor group. HUVECs apoptosis were assessed by fluorescent staining with Hoechst33258 and flow cytometry with Annexin V-PI kit staining. Western blotting was used to detect the protein expression of PKB, p-PKB （Threonine308） and p-PKB （Serine473）. RESULTS The apoptotic index （AI） of high glucose group （ 14.07 ± s 0.25） % increased significantly compared to that of control group （P 〈 0.01）, but the AI of ibrolipim 8 μmol·L^-1 group （5.42 ± 0.13）% decreased significantly compared to that of high glucose group （P 〈 0.01）, and the AI of inhibiter group （12.00 ± 0.10） % increased compared to that of ibrolipim group （P 〈 0.01 ） . Low concentration ibrolipim inhibited high glucose-induced apoptosis of cultured HUVECs＇during early time, with subsequent increase in PKB phosphorylation （P 〈 0.01 ）. Blocking PKB phosphorylation significantly abolished the anti-apoptotic effect of ibrolipim. CONCLUSION Ibrolipim at low concentration can inhibit high glucose-induced apoptosis of cultured HUVECs, which may be related to PKB phosphorylation.

关 键 词：糖尿病 动脉粥样硬化 葡萄糖 蛋白质丝氨酸苏氨酸激酶 内皮细胞 凋亡 艾溴利平 

分 类 号：R972.6[医药卫生—药品]