中国病理生理杂志2019，Vol.35Issue(2)：332-339,8.DOI:10.3969/j.issn.1000-4718.2019.02.023
内质网应激诱导的自噬对肝细胞凋亡的影响
Effect of endoplasmic reticulum stress-induced autophagy on hepatocyte apoptosis
摘要
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.关键词
肝细胞/内质网应激/细胞凋亡/自噬/雷帕霉素Key words
Hepatocytes/Endoplasmic reticulum stress/Apoptosis/Autophagy/Rapamycin分类
医药卫生引用本文复制引用
郑璐,谢汝佳,韩冰,汤雷,田甜,蔡爽,余蕾,马子华,杨婷,杨勤..内质网应激诱导的自噬对肝细胞凋亡的影响[J].中国病理生理杂志,2019,35(2):332-339,8.基金项目
国家自然科学基金资助项目(No.81560105) （No.81560105）
贵州省科技计划项目(黔科合LH字[2014]7074) （黔科合LH字[2014]7074）
贵州省科技合作计划项目(黔科合LH字[2015]7347) （黔科合LH字[2015]7347）
