自噬参与高糖缺氧诱导的人脐静脉内皮损伤机制的实验研究

目的 观察高糖缺氧状态人脐静脉内皮细胞（HUVECs）自噬水平的变化，探讨自噬在这一过程中所起到的作用。 方法 通过体外细胞培养的方法培养HUVECs并分为6组，即正常对照组、高糖组、正常细胞缺氧组、高糖缺氧组、高糖缺氧+雷帕霉素组、高糖缺氧+3-甲基腺嘌呤（3-MA）组。各组经过相应处理后，通过流式细胞仪、Caspase-3活性试剂盒检测凋亡率、Western blot检测自噬相关蛋白（LC-3、Beclin-1、Atg-5和STSQM1）的表达情况。 结果 与正常缺氧组相比，高糖缺氧组的自噬相关蛋白LC-3B、Beclin-1和Atg-5表达降低，STSQM1表达升高（均P<0.05），使用雷帕霉素可以上调高糖缺氧组自噬水平并且降低细胞凋亡率，加入自噬抑制剂3-MA使高糖缺氧组自噬下调，细胞凋亡率上升（均P<0.05）。 结论 高糖缺氧状态下HUVECs自噬功能受损，上调自噬可以降低凋亡率，发挥保护细胞的作用。

Effects of autophagy on high glucose/hypoxia induced human umbilical vein endothelial cells (HUVECs) injury

AIM To investigate the effect of autophagy on human umbilical vein endothelial cells (HUVECs) in high concentration glucose and hypoxia contexts. METHODS Cultured HUVECs were divided into six groups: control group, high glucose group, control hypoxia group, high glucose hypoxia group, high glucose hypoxia plus Rapamycin group, and high glucose hypoxia plus 3-MA group. Apoptosis was evaluated by Annexin V-FITC/PI apoptosis detection kit, expressions of LC-3 and STSQM1 were analyzed by Western blot and LC-3 expression was detected by immunofluorescence. RESULTS Apoptosis rate was highly increased in high glucose and high glucose hypoxia group compared with that in control group. However, rapamycin decreased the apoptosis of HUVECs and autophagy inhibitor 3-MA increased the apoptosis of HUVECs induced by high glucose and hypoxia P<0.05). CONCLUSION HUVECs' functional autophagy is undermined in high glucose and hypoxia contexts and the up-regulation of autophagy decreases the apoptosis rate, thus exerting the protective effect of HUVECs.