主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (11): 933-939.doi: 10.3969/j.issn.1006-2157.2017.11.011

• 科技之窗 • 上一篇    下一篇

丹参酮ⅡA通过PI3K/Akt/mTOR信号通路调控自噬抗内皮细胞氧化应激损伤研究*

曹慧敏1,宋囡1,张妮1,杨关林1,陈文娜1,张哲2,贾连群1#   

  1. 1 辽宁中医药大学中医脏象理论及应用教育部重点实验室 辽宁省中医转化医学研究中心辽宁 110847;
    2 辽宁中医药大学附属医院
  • 收稿日期:2017-05-22 出版日期:2017-11-10 发布日期:2017-11-10
  • 通讯作者: 贾连群,女,博士,教授,主要研究方向:中西医结合防治心血管疾病,E-mail:jlq-8@163.com
  • 作者简介:曹慧敏,女,在读硕士生
  • 基金资助:
    国家自然科学基金青年基金项目(No.81300229),辽宁省自然科学基金项目(No.2015020394),辽宁省高等学校优秀人才支持计划(No.LR2015041),辽宁中医药大学2015年度大学生创新创业训练计划项目(No.201510162000066)

Regulation on autophagy with tanshinone ⅡA for anti-oxidative stress damage of endothelial cells through PI3K/Akt/mTOR pathway*

CAO Huimin1, SONG Nan1, ZHANG Ni1, YANG Guanlin1, CHEN Wenna1, ZHANG Zhe2, JIA Lianqun1#   

  1. 1 Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Translational Medicine Research Center of traditional Chinese medicine in Liaoning Province, Liaoning University of Traditional Chinese Medicine, Liaoning 110847, China;
    2 Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Liaoning 110032, China
  • Received:2017-05-22 Online:2017-11-10 Published:2017-11-10
  • Supported by:
    National Natural Science Foundation for Young Scientists of China (No. 81300229), Natural Science Foundation of Liaoning Province (No. 2015020394), Support Programme for Talents in Colleges and Universities of Liaoning Province (No. LR2015041)

摘要: 目的 基于PI3K/Akt/mTOR 自噬信号通路探讨丹参酮ⅡA对氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导内皮细胞氧化应激损伤的保护作用及机制。方法 体外培养EA.hy926细胞,随机分为正常组、模型组、丹参酮ⅡA组、LY294002组、ox-LDL加丹参酮ⅡA组、ox-LDL加丹参酮ⅡA加LY294002组。用比色法检测细胞氧化应激损伤丙二醛(MDA)含量及超氧化物歧化酶(SOD)活力,利用倒置荧光显微镜及Naolive 3D cell explorer实时无标记3D显微成像系统检测自噬发生,同时应用免疫印迹(Western Blotting)检测自噬相关蛋白以及PI3K/Akt/mTOR通路蛋白的表达情况。结果 与正常组相比,模型组MDA含量增高,SOD活力降低,微管相关蛋白1轻链3(LC3)I/II蛋白含量增多(P<0.01);丹参酮ⅡA干预后细胞中MDA含量降低,SOD活力增高,LC3-I/LC3-II蛋白含量增多(P<0.01)。与正常组相比,模型组PI3K、p-Akt、p-mTOR蛋白表达水平明显减少(P<0.05或P<0.01);与模型组相比,丹参酮ⅡA干预后细胞中PI3K、p-Akt、p-mTOR蛋白表达水平明显减少(P<0.05或P<0.01);与ox-LDL加丹参酮ⅡA组相比,加入PI3K的抑制剂LY294002后细胞中的LC3-I/LC3-II蛋白含量减少,自噬水平减少(P<0.01)。结论 丹参酮ⅡA可能通过调控PI3K/Akt/mTOR信号通路促进自噬,对ox-LDL诱导的EA. Hy926细胞氧化应激损伤起到保护作用,进而防治动脉粥样硬化。

关键词: 丹参酮ⅡA, ox-LDL, 动脉粥样硬化, 氧化应激, 自噬

Abstract: Objective To investigate the protective and mechanism of tanshinone ⅡA (TSⅡA) in oxidative stress damage of endothelial cells induced by oxidized low-density lipoprotein (ox-LDL) based on PI3K/Akt/mTOR autophagy pathway. Methods EA. hy926 cells were cultured in vitro, and then randomly divided into normal group, model group, ox-LDL+TSⅡA group, ox-LDL+TSⅡA+LY294002 group, TSⅡA group and LY294002 group. The content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were detected by using chromatoptometry, autophagy status was observed by using FITC-LC3 fluorescence microscope and Naolive 3D Cell Explorer Real-time Microscopic Imaging System, and meanwhile, expressions of autophagy-related proteins and PI3K/Akt/mTOR pathway proteins were detected by using Western Blotting. Results Compared with normal group, MDA content increased, SOD activity decreased and content of LC3-I/LC3-II protein increased in model group (P<0.01), and after TSⅡA intervention, MDA content decreased, SOD activity increased and content of LC3-I/LC3-II protein increased (P<0.01). Compared with normal group, protein expressions of PI3K, p-Akt and p-mTOR decreased significantly in model group (P<0.05 or P<0.01). Compared with model group, protein expressions of PI3K, p-Akt and p-mTOR decreased significantly after TSⅡA intervention (P<0.05 or P<0.01). Compared with ox-LDL+TSⅡA group, content of LC3-I/LC3-II protein decreased and autophagy level decreased (P<0.01) after applying LY294002, an inhibitor of PI3K. Conclusion Tanshinone ⅡA may improve autophagy through regulating PI3K/Akt/mTOR pathway to relieve the oxidative stress damage of EA. Hy926 cells and prevent atherosclerosis.

Key words: tanshinone ⅡA, oxidized low-density lipoprotein, atherosclerosis, oxidative stress, autophagy

中图分类号: 

  • R285.5