高糖抑制PI3K/AKT通路诱导人冠状动脉内皮细胞凋亡的研究  

High glucose induces human endothelial cell apoptosis through inhibiting PI3K /AKT pathway

作　　者：崔丽[1] 李纪明[1] 罗轶玮[1] 方明[1] 刘学波[1] 李莹[1] 

机构地区：[1]同济大学附属东方医院心内科,上海200120

出　　处：《同济大学学报（医学版）》2014年第1期30-34,共5页Journal of Tongji University(Medical Science)

基　　金：上海市科委攻关项目(10411968000);上海市自然科学基金(10ZR1406600)

摘　　要：目的探讨高糖对人冠状动脉内皮细胞凋亡及微颗粒释放的影响及机制。方法根据细胞培养基中葡萄糖浓度不同分为3组:正常对照组(5.5 mmol/L),中糖组(17.6 mmol/L),高糖组(33.3 mmol/L),另设甘露醇对照组(20.0 mmol/L)。通过Hoechst 33258荧光染色观察凋亡形态学变化,应用流式细胞术检测内皮微颗粒细胞凋亡率及微颗粒,ELISA法检测人冠状动脉内皮细胞的凋亡相关蛋白Bad、Bax的表达以及PI3K、p-Akt蛋白的表达。结果随着葡萄糖浓度增加,人冠状动脉内皮细胞微颗粒的释放及细胞凋亡率逐渐增加,高糖组最高(P<0.01);人冠状动脉内皮细胞分泌Bad、Bax逐渐增多,高糖组最高(P<0.01);人冠状动脉内皮细胞分泌PI3K、pAkt逐渐减少,高糖组减少最明显(P<0.01)。与甘露醇对照组比较,正常对照组微颗粒的释放,细胞凋亡率,分泌Bad、Bax、PI3K、p-Akt差异均无统计学意义(P>0.05)。结论高糖可以促进人冠状动脉内皮细胞凋亡及微颗粒的释放,其机制可能同PI3K/AKT途径相关。Objective To investigate the effect of high glucose on human coronary endothelial cell （HCAEC） apoptosis and its mechanism. Methods Cultured HCAECs were treated with normal （5.5 mmol/L）, medium （17.6 mmol/L） and high concentration of glucose （33.3 mmol/L）, or mannitol （20.0 mmol/L）, respectively. Hoechst 33258 fluorescence staining was used to observe the morphological changes of apoptosis. The absolute number of endothelial microparticles and the apoptotic cells percentage were identified by flow cytometer; the expression of Bad, Bax, PI3K and p- Akt in cell lysates were detected by ELISA. Results HCAECs microparticles and apoptotic cells were increased with the increasing glucose concentrations and those in high glucose group increased most （ P 〈 0.01 ）. Bad and Bax secreted by HCAECs increased with the increasing of glucose concentrationsand those in high glucose group increased most （P 〈 0. 01 ）. PI3K and p-Akt secreted by HCAECs secreted were decreased and those in high glucose group decreased most （P 〈0. 01）. There were no significant differences in HCAECs microparticles, apoptotic cells, Bad, Bax, PI3K and p-Akt expressions between control group and mannitol group （P 〉 0.05 ）. Conclusion High glucose induces HCAECs apoptosis and EMPs release, in which PI3K/AKT pathway may be involved.

关 键 词：人冠状动脉内皮细胞 凋亡 高糖 内皮微颗粒 PBK AKT 

分 类 号：R543.3[医药卫生—心血管疾病]