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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 role of gas in autophagy induced by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were pretreated with different concentrations of Gas (5, 10, 20 and 50 μmol / L) for 1 h and then incubated with 1 mg / L LPS for 12 h. Total protein and mRNA were extracted respectively. Western blotting and RT- Beclin-1, ATG5 and LC3 were detected by flow cytometry. The expression of autophagosomes was detected by Dansyl-cadaverine (MDC) staining. The survival rate of HUVECs was detected by MTT assay. Results LPS induced autophagy in HUVECs. After pretreatment with Gas, the level of autophagy was significantly reduced and in a dose-dependent manner. At the same time, the survival rate of cells was significantly increased. Conclusion LPS can induce autophagy in HUVECs. Gas can increase the survival rate of HUVECs by inhibiting autophagy induced by LPS.