天然产物研究与开发 ›› 2019, Vol. 31 ›› Issue (9): 1537-1542.doi: 10.16333/j.1001-6880.2019.9.008

• 研究论文 • 上一篇    下一篇

细叶远志皂苷在Aβ23-35诱导SH-SY5Y细胞氧化损伤中的作用及机制研究

王 琳,金桂芳,余河汉,李 巧,杨 红*   

  1. 广东药科大学生命科学与生物制药学院,广州 510006
  • 出版日期:2019-10-09 发布日期:2019-10-23
  • 基金资助:

    广州市科技计划(201707010409);广州市特色创新类项目(2017KTSCX105)

Effects of tenuifolin against oxidative damage induced by Aβ25-35 in SH-SY5Y cells and underlying mechanisms

WANG Lin,JIN Gui-fang,YU He-han,LI Qiao,YANG Hong*   

  1. College of Life Sciences and Biopharmaceuticals of Guangdong Pharmaceutical University,Guangzhou 510006,China
  • Online:2019-10-09 Published:2019-10-23

摘要: 为研究细叶远志皂苷(tenuifolin,TEN)在Aβ25-35诱导SH-SY5Y细胞氧化损伤中的作用,并探讨其作用机制。建立Aβ25-35诱导的细胞损伤模型,细叶远志皂苷以及自噬抑制剂3-MA进行干预,显微镜观察细胞形态变化,试剂盒检测细胞氧化应激水平,RT-qPCR和Western blot检测细叶远志皂苷以及自噬抑制剂干预前后Beclin-1、LC3、mTOR、AMPK和ULK1 mRNA及蛋白水平变化。结果发现,TEN改善Aβ25-35诱导的SH-SY5Y细胞形态损伤和细胞活力下降;降低ROS和MDA浓度,并提高SOD、GSH-Px及过氧化氢酶的活性;增加AMPK和ULK1 的表达,减少mTOR的表达及增加Beclin-1和LC3-II/I的表达水平。而加入3-MA会拮抗TEN的作用。总之,TEN可能通过调控AMPK/mTOR/ULK1通路,增加Beclin-1及LC3-II/I蛋白水平激活自噬,进而改善Aβ25-35诱导的细胞形态损伤和细胞活力下降,提高细胞抗氧化应激能力,发挥神经保护作用。

关键词: 细叶远志皂苷, 阿尔兹海默症, &beta, -淀粉样蛋白25-35, 细胞自噬, 氧化应激

Abstract: In order to investigate the effects of tenuifolin (TEN) in the induction of SH-SY5Y cell oxidative damage induced by Aβ25-35 and the underlying mechanisms.Aβ25-35-induced cell injury models were established.Interferon TEN and autophagy inhibitor 3-MA were intervened.The morphological changes were observed by microscopy.The oxidative stress levels were detected by kits.RT-PCR and Western blot were used to detect the changes in Beclin-1,LC3,mTOR,AMPK and ULK1 mRNA and protein levels before and after TEN and 3-MA.It was found that TEN improved the morphological damage and cell viability of SH-SY5Y cells induced by Aβ25-35.Compared with Aβ25-35 group,TEN decreased the concentration of ROS and MDA,and increased the activity of SOD,GSH-Px and CAT.The expression levels of LC3-II/I and Beclin-1 increased in Aβ25-35 group,and it's further increased in TEN group.And TEN increased the expression of AMPK and ULK1,decreased the expression of mTOR.The addition of 3-MA antagonizes the action of TEN.However,3-MA reversed the effects of TEN.In conclusion,it can be proved thatTEN may activate autophagy by regulating AMPK/ mTOR/ ULK1 pathway to inhibit the cell oxidative damage induced by Aβ25-35,and play a neuroprotective role.

Key words: tenuifolin, Alzheimer's disease, &beta, -amyloid protein 25-35, autophagy, oxidative stress

中图分类号: 

R285