Effect of endoplasmic reticulum stress-induced autophagy on hepatocyte apoptosis
10.3969/j.issn.1000-4718.2019.02.023
- VernacularTitle:内质网应激诱导的自噬对肝细胞凋亡的影响
- Author:
Lu ZHENG
1
;
Bing HAN
;
Lei TANG
;
Tian TIAN
;
Shuang CAI
;
Lei YU
;
Zi-Hua MA
;
Ting YANG
;
Qin YANG
;
Ru-Jia XIE
Author Information
1. 贵州医科大学病理生理学教研室
- Keywords:
Hepatocytes;
Endoplasmic reticulum stress;
Apoptosis;
Autophagy;
Rapamycin
- From:
Chinese Journal of Pathophysiology
2019;35(2):332-339
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To observe the changes of autophagy-related indexes during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT) and its effect on apoptosis in human normal hepatocytes.METHODS:LO2 cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS.The expression of glucose-regulated protein 78 (GRP78) , protein kinase R-like endoplasmic reticulum kinase (PERK) , activating transcription factor 4 (ATF4) , C/EBP homologous protein (CHOP) , autophagy-related gene 12 (Atg12) , autophagy-related gene 5 (Atg5) and microtubule-associated protein 1 light chain 3 (LC3) at mRNA and protein levels was determined by real-time PCR and Western blot.The apoptosis was analyzed by flow cytometry.The formation of autophagosomes was observed under transmission electron microscope.After the LO2 cells were pretreated with rapamycin at 400 nmol/L for 1 h and treated with DTT at 2.0mmol/L for 24 h, the effect of rapamycin pretreatment on the apoptosis was analyzed by flow cytometry.RESULTS:After treatment with DTT at 2.0 mmol/L for 6, 12 and 24 h, the mRNA and protein levels of GRP78, PERK, ATF4, CHOP, Atg12, Atg5 and LC3 in the LO2 cells were significantly higher than those in 0 h group (P<0.05).At the same time, the ratio of LC3Ⅱ/LC3Ⅰwas also increased after DTT treatment (P<0.05).Observation under transmission electron microscope showed that autophagosomes were found in the LO2 cells treated with DTT for 6, 12 and 24 h.After DTT treatment for 6, 12 and 24 h, the apoptosis rate of LO2 cells was significantly higher than that in DTT 0 h group, while the apoptosis induced by DTT was significantly decreased after rapamycin pretreatment (P<0.05).CONCLUSION:ERS induces autophagy and rapamycin pretreatment alleviates the apoptosis of LO2 cells to some extent.
