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天麻素抑制脂多糖诱导的人脐静脉内皮细胞的自噬
Gastrodin inhibits lipopolysaccharide-induced autophagy in human umbilical vein endothelial cells
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。
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.
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
Gastrodin / Lipopolysaccharide / Autophagy / Monodansylcadaverine / Western blotting / RT-PCR / Human umbilical vein endothelial cell
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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
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