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 1 h before 1 mg/L LPS stimulation, after 12h incubation. The protein and mRNA expression of Beelin- 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.