奋乃静预处理缓解衣霉素诱发的细胞内质网应激

Pretreatment with perphenazine alleviated endoplasmic reticulum stress induced by tunicamycin

目的研究奋乃静预处理对衣霉素诱发细胞内质网应激的影响及机制。方法用Western blot、real time-PCR、Hoechst33342染色及Filipin染色技术检测奋乃静预处理后MLE12细胞对衣霉素诱发内质网应激的改变,检查奋乃静预处理后自噬相关分子、细胞分化标志性分子表达的变化,评估细胞自噬水平以及溶酶体功能状态。结果相比未预处理细胞,奋乃静预处理后缓解衣霉素诱发的内质网应激,凋亡细胞减少。低剂量奋乃静可使MLE12细胞自噬启动增强,内体溶酶体功能受到影响,细胞分化标志SPB、SPC表达上升。结论奋乃静预处理增强MLE12细胞自噬,提升细胞对抗衣霉素诱发的内质网应激能力,促进其向Ⅱ型肺泡上皮细胞分化。

Objective To analyze the effects and biological mechanism of perphenazine preconditioning on endoplasmic reticulum stress induced by tunicamycin.Methods Western blot,Real time-PCR,Hoechst33342 staining and Filipin staining were used to detect the changes of endoplasmic reticulum stress level in tunicamycin-induced perphenazine preconditioning MLE12 cells,examine the changes in the expression of autophagy-related molecules and marker molecules of cell differentiation and assess of autophagy levels and lysosomal functional status after perphenazine treatment.Results Compared with untreated cells,perphenazine pretreatment relieved tunicamycin-induced endoplasmic reticulum stress,and decreased apoptotic cells.Low-dose perphenazine enhanced the autophagy initiation of MLE12 cells,affected the lysosomal function of endosomes,and increased the expression level of SPB and SPC in cell differentiation markers.Conclusion Perphenazine preconditioning enhances autophagy and ability of MLE12 cells to antagonize endoplasmic reticulum stress induced by tunicamycin,and promotes its differentiation into typeⅡalveolar epithelial cells.