解剖学报 ›› 2014, Vol. 45 ›› Issue (6): 824-829.doi: 10.3969/j.issn.0529-1356.2014.06.016

• 组织学胚胎学发育生物学 • 上一篇    下一篇

天麻素抑制脂多糖诱导的人脐静脉内皮细胞的自噬

解亚男1 杨萍1 文飞1 孙林2 陈媛丽1* 陆地1*   

  1. 1.昆明医科大学人体解剖学与组织学胚胎学系, 昆明 650500; 2. 昆明医科大学第二附属医院心内科, 昆明 650101
  • 收稿日期:2014-03-31 修回日期:2014-06-20 出版日期:2012-12-06 发布日期:2014-12-06
  • 通讯作者: 陈媛,陆地 E-mail:ludi20040609@126.com
  • 基金资助:

    昆明医科大学研究生创新基金项目;国家自然科学基金资助项目;云南省应用基础研究重点项目

Gastrodin inhibits lipopolysaccharide-induced autophagy in human umbilical vein endothelial cells

XIE Ya-nan1 YANG Ping1 WEN Fei1 SUN Lin2 CHEN Yuan-li 1* LU Di 1*   

  1. 1. Department of Human Anatomy and Histological Embryology,Kunming Medical University, Kunming 650500,China;2. Department of Cardiology, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, China
  • Received:2014-03-31 Revised:2014-06-20 Online:2012-12-06 Published:2014-12-06
  • Contact: CHEN Yuan-li , LU Di E-mail:ludi20040609@126.com

摘要:

目的 探讨天麻素(Gas)在脂多糖 (LPS) 诱导的人脐静脉内皮细胞 ( HUVECs) 自噬中的作用。方法 用不同浓度的Gas(5、10、20、50μmol/L) 预处理 HUVECs 1h,再加入1mg/L LPS共培养 12 h后,分别提取细胞总蛋白及mRNA,采用 Western blotting 及 RT-PCR 检测 Beclin-1、ATG5 及 LC3的表达变化;采用丹酰尸胺( MDC) 染色检测细胞中自噬小体(autophagosome)的表达变化;采用MTT检测 HUVECs 生存率的变化。 结果 LPS 诱导 HUVECs 产生自噬;Gas 预处理后自噬水平明显降低且呈剂量依赖性;与此同时细胞的生存率明显增加。 结论 LPS诱导 HUVECs 产生自噬;Gas 通过抑制由LPS 诱导产生的自噬,提高 HUVECs 的生存率。

关键词: 天麻素, 脂多糖, 自噬, 丹酰尸胺, 免疫印迹法, 反转录-聚合酶链反应, 人脐静脉内皮细胞

Abstract:

Objective To investigate the regulation mechanism of gastrodin (Gas) on atutophagy of human umbilical vein endothelial cells(HUVECs)stimulated with lipopolysaccharide (LPS). Methods Gas at 5, 10, 20 and 50μmol/L was added 1h before 1mg/L LPS stimulation, after 12h incubation. The protein and mRNA expression of Beclin-1, ATG5 and LC3 were detected by Western blottingting and RT-PCR respectively. The autophagosome was observed under fluorescent microscope by monodansylcadaverine (MDC) staining. Cell viability rate was measured by MTT analysis. Results Autophagy was induced in HUVECs by LPS-stimulation. The protein and mRNA expression of Beclin-1, ATG5 and LC3 were significantly increased,which was in a time-dependent manner. Compared with LPS-stimulated group, pertreatment with Gas inhibited the expression of autophagy in dose-dependent manner and the cell viability was effectively increased. Conclusion Autophagy was induced in HUVECs by LPS-stimulation. Gas may increase the viability rate of HUVECs through inhibiting the expression of autophagy.

Key words: Gastrodin, Lipopolysaccharide, Autophagy, Monodansylcadaverine, Western blotting, RT-PCR, Human umbilical vein endothelial cell