目的 研究小檗碱对波动性高糖所致人冠状动脉内皮细胞（HCAECs）凋亡的影响。方法 分离、鉴定并培养HCAECs，将HCAECs分为5组：正常葡萄糖组（5.5 mmol/L葡萄糖）、持续性高糖组（25 mmol/L葡萄糖）、波动性高糖组（5.5、25 mmol/L葡萄糖每24小时波动1次）、波动性高渗环境（不加入葡萄糖，加入与波动性高糖组相同浓度的甘露醇，保持与波动性高糖组相同的波动性高渗环境）、波动性高糖+小檗碱干预组（5.5、25 mmol/L葡萄糖+50 μmol/L小檗碱每24小时波动1次），各组细胞每24小时换液1次，共培养7 d后进行后期实验。检测各组HCAECs的细胞活力及凋亡情况，利用实时荧光定量（qRT-PCR）及Western blotting技术检测细胞凋亡相关蛋白Caspase-3表达量。结果 与持续性高糖组相比，波动性高糖组HCAECs凋亡更显著（P<0.05），波动性高糖组Caspase-3蛋白及mRNA表达水平明显增加（P<0.05）；但波动性高糖+小檗碱干预组较波动性高糖组HCAECs凋亡明显减弱（P<0.05），小檗碱可以明显下调波动性高糖诱导的Caspase-3表达（P<0.05）。结论 波动性高糖较持续性高糖更易降低HCAECs活力，促进HCAECs凋亡，但小檗碱显著减弱波动性高糖环境下HCAECs凋亡，对HCAECs具有明显的保护作用。

Objective To investigate the effect of berberine on the apoptosis of human coronary artery endothelial cells (HCAECs) induced by intermittent high glucose in vitro. Methods HCAECs were isolated, identified, cultured, and divided into five groups:Group A was the normal glucose group (5.5 mmol/L glucose, NG); Group B was the persistent high glucose group (25 mmol/L glucose, PHG); Group C was the intermittent high glucose group (5.5 mmol/L and 25 mmol/L glucose fluctuated every 24 h, IHG); Group D was in a volatile hyperosmotic environment (5.5 mmol/L and 25 mmol/L mannitol alternating every 24 h to maintain the same osmotic pressure as IHG, OC); Group E was the fluctuation of hyperglycemia + berberine intervention group (5.5 mmol/L and 25 mmol/L glucose + 50 μmol/L berberine fluctuated every 24 h, IHG + BBR). The cells of each group were changed every 24 h, and the culture was stopped after 7 d, the cell viability and apoptosis were detected in each group. The expression of the apoptosis related protein Caspase-3 was determined by qRT-PCR and Western blotting. Results Compared with the persistent high glucose group, the apoptosis of HCAECs in the intermittent high glucose group was more significant (P < 0.05), but the apoptosis of HCAECs in the fluctuation of hyperglycemia + berberine intervention group was more significantly reduced than that in the intermittent high glucose group (P < 0.05). Berberine significantly reduced the expression of Caspase-3 induced by intermittent high glucose (P < 0.05) Conclusion Intermittent high glucose decreased the activity of HCAECs more easily than the persistent high glucose,and promoted the apoptosis of HCAECs. But berberine significantly reduced the apoptosis of HCAECs under intermittent high glucose, which had a significant protective effect on HCAECs.

R085.5

江苏省“六大人才高峰”资助项目（2014-WSN-045）