白藜芦醇通过诱导ROS及活化AMPK促进Hep-2细胞自噬作用

摘要

Objective To explore effect of Resveratrol on autophagy of Hep‐2 Cell and analysis of relative mecha‐nisms . Methods We apply western blotting and real time‐PCR to detect the protein levels .Intracellular reactive oxygen species (ROS) level was assessed by DCFH‐DA staining .All data are analyzed by using SPSS 13 software . Results Resveratrol promote Hep‐2 cells autophagy ,with 40 μM and 6 h in the most significant .It can increase the ratio of LC3 Ⅱ and LC3 I ,promoting the protein expression of beclin‐1 and the number of autophagic vacuoles and autophagosomes were also increased significantly .Mechanism study found that the effect of resveratrol promo‐ting Hep‐2 cells autophagy related to ROS increased .After resveratrol treatment ,the ROS in Hep‐2 cells increase about 6 times .Further research found that ROS induces Hep‐2 cells autophagy through the activation of AM PK‐mTOR pathway .The promotion role of resveratrol on apoptosis in Hep‐2 cells was weakened significantly with u‐sing 3MA and CQ inhibiting autophagy . Conclusions Resveratrol can promote Hep‐2 cells autophagy through AMPK‐mTOR pathway ,and this pathway activated by ROS .%目的：探讨白藜芦醇通过诱导 ROS及活化 AMPK 促进 Hep‐2细胞自噬的可能机制。方法采用40μM 浓度白藜芦醇复合培养液作用于 Hep‐2细胞6 h后，western blot分别分析蛋白水平，DCFH‐DA 染色法分析细胞内活性氧水平。结果白藜芦醇促 Hep‐2细胞自噬作用与其促活性氧增多有关，经白藜芦醇处理后，Hep‐2细胞内活性氧增加约6倍，进一步研究发现，活性氧通过激活 AMPK‐mTOR途径而促进 Hep‐2细胞自噬。结论白藜芦醇诱导 Hep‐2细胞自噬的机制可能与通过活性氧激活 AMPK‐mTOR途径促进 Hep‐2细胞自噬有关。