姜黄素对过氧化氢诱导内皮细胞凋亡及衰老的影响

目的 探讨姜黄素对过氧化氢(H2O2)诱导人脐静脉内皮细胞(HUVECs)自噬、凋亡及衰老的影响.方法 体外培养HUVECs,分为对照组(只给予培养基),模型组(给予60 μmol/L H2O2),姜黄素组(给予60μmol/L H2O2+ 10 μmol/L姜黄素),3-MA组[(给予60 μmol/L H2O2+ 10μmol/L姜黄素+1 mmol/L3-甲基腺嘌呤(3-Methyladenine)],培养4d后,DCFH-DA法检测细胞培养液中活性氧(ROS)的含量,流式细胞仪检测细胞凋亡水平,衰老相关β-半乳糖苷酶(SA-β-gal)染色检测细胞衰老程度,Western blot技术检测自噬相关蛋白LC3和p62的表达水平.结果 与对照组相比,模型组ROS含量、凋亡率、SA-β-gal阳性率及p62水平升高(均P＜0.05),而LC3-Ⅱ/Ⅰ比值差异无统计学意义(P＞0.05).与模型组比,姜黄素组ROS含量、凋亡率及SA-β-gal阳性率及p62水平降低(均P＜0.05),但LC3-Ⅱ/Ⅰ比值升高(P＜0.05).与姜黄素组比较,3-MA组ROS含量、凋亡率、SA-β-gal阳性率及p62水平升高(均P＜0.05),但LC3-Ⅱ/Ⅰ比值降低(P＜0.05).结论 姜黄素可减轻H2O2诱导内皮细胞凋亡及衰老,其机制与提高内皮细胞自噬有关.

Curcumin attenuates H2O2-induced apoptosis and premature senescence in endothelial cells via autophagy up-regulation

Objective To investigate the effects of curcumin (Cur) on autophagy,apoptosis and premature se nescence induced by H2 O2 in human umbilical vein endothelial cells (HUVECs).Methods HUVECs were cultured in vitro and randomized into control group (treated with culture medium),model group (treated with 60 μmoL/L H2O2),Cur group (treated with 60 μmol/L H2O2 and 10 μmol/L Cur) and 3-MA group (treated with 60 μmol/L H2O2,10 μmol/L Cur and 1 mmol/L 3-Methyladenine).After 4 days of culture,DCFH-DA was used to measure the level of reactive oxygen species (ROS) in endothelial cells.Cells apoptosis were detected by flow cytometry.SA-β3-gal staining was used to identify cells aging status.Western blot was conducted to detect the expression of autophagy marking protein LC3 and p62.Results Compared with control group,the level of ROS,apoptosis rate,the number of SA-β-gal positive cells and the expression of p62 protein were significantly increased (P ＜ 0.05) in model group,while the expression of LC3-Ⅱ / Ⅰ protein showed no significant difference (P ＞ 0.05).Compared with model group,the level of ROS,apoptosis rate,the number of SA-β-gal positive cells and the expression of p62 protein were significantly reduced (P ＜ 0.05) in Cur group,and the expression of LC3-Ⅱ / Ⅰ protein was significantly increased (P ＜ 0.05).All the effects of curcumin mentioned above were inhibited by 3-MA,the autophagy inhibitor (P ＜ 0.05).Conclusion Curcumin can attenuate H2 O2-induced apoptosis and premature senescence in endothelial cells via autophagy up-regulation.