PMID- 31416082 OWN - NLM STAT- MEDLINE DCOM- 20200922 LR - 20211204 IS - 1423-0356 (Electronic) IS - 0025-7931 (Linking) VI - 98 IP - 4 DP - 2019 TI - Cigarette Smoke Extract Promotes Human Lung Myofibroblast Differentiation by the Induction of Endoplasmic Reticulum Stress. PG - 347-356 LID - 10.1159/000502099 [doi] AB - BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a progressive and lethal fibrotic lung disease with an unknown aetiology. Persistent myofibroblast differentiation is a prominent feature of IPF. Cigarette smoking is a risk factor for IPF and an indicator of poor prognosis. Cigarette smoking induces endoplasmic reticulum (ER) stress, and it has been shown that ER stress promotes fibroblast-to-myofibroblast differentiation in lung fibrosis. In this study, we investigated whether cigarette smoke extract (CSE) promotes lung myofibroblast differentiation via the induction of ER stress. OBJECTIVES: Our study concentrates on exploring the relationship between smoking and ER stress in the differentiation of lung fibroblasts to myofibroblasts. METHODS: Human embryonic lung fibroblasts (MRC-5 fibroblasts) were stimulated with various doses of CSE. Levels of alpha-smooth muscle actin (alpha-SMA) protein were evaluated by immunofluorescence and western blot analyses. ER stress was induced by thapsigargin (TG) and inhibited by 4-phenyl butyric acid (4-PBA). Protein levels of glucose-regulated protein-78 (GRP78), inositol-requiring enzyme 1 (IRE1), X box-binding protein-1 (XBP-1) and activating transcription factor 6 (ATF6) were determined by western blotting. GRP78 siRNA was transfected into MRC-5 cells using Lipofectamine RNAiMAX Reagent. RESULTS: CSE at a concentration of 1.0% significantly increased alpha-SMA expression in MRC-5 cells. There was no significant cell apoptosis after cells were exposed to CSE. CSE treatment significantly increased the expression of GRP78, IRE1, XBP-1 and ATF6 at the protein level at 48 h. Pretreatment with TG enhanced, whereas pretreatment with 4-PBA inhibited, the CSE-induced expression of alpha-SMA, GRP78 and XBP-1. Furthermore, knockdown of GRP78 blocked alpha-SMA expression in MRC-5 cells exposed to CSE. CONCLUSION: Our data suggested that CSE promotes lung fibroblast-to-myofibroblast differentiation by the induction of ER stress. CI - (c) 2019 S. Karger AG, Basel. FAU - Song, Min AU - Song M AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China. FAU - Peng, Hong AU - Peng H AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China, penghong66@csu.edu.cn. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China, penghong66@csu.edu.cn. FAU - Guo, Wei AU - Guo W AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China. FAU - Luo, Man AU - Luo M AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China. FAU - Duan, Wang AU - Duan W AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China. FAU - Chen, Ping AU - Chen P AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Respiratory Disease Diagnosis and Treatment Center of the Central South University, Changsha, China. FAU - Zhou, Yong AU - Zhou Y AD - Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Respiratory Disease Research Institute of Hunan Province, Changsha, China. AD - Division of Pulmonary, Department of Medicine, Allergy and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA. LA - eng PT - Journal Article DEP - 20190815 PL - Switzerland TA - Respiration JT - Respiration; international review of thoracic diseases JID - 0137356 RN - 0 (Endoplasmic Reticulum Chaperone BiP) RN - 0 (HSPA5 protein, human) RN - 67526-95-8 (Thapsigargin) SB - IM MH - Cell Differentiation/*drug effects MH - Cell Line MH - Cigarette Smoking/*adverse effects MH - Endoplasmic Reticulum Chaperone BiP MH - *Endoplasmic Reticulum Stress MH - Fibroblasts/*drug effects MH - Humans MH - Idiopathic Pulmonary Fibrosis/*etiology MH - Lung/cytology MH - Thapsigargin OTO - NOTNLM OT - Cigarette smoking OT - Endoplasmic reticulum stress OT - Fibroblast OT - Idiopathic pulmonary fibrosis OT - Myofibroblast differentiation EDAT- 2019/08/16 06:00 MHDA- 2020/09/23 06:00 CRDT- 2019/08/16 06:00 PHST- 2018/11/27 00:00 [received] PHST- 2019/07/10 00:00 [accepted] PHST- 2019/08/16 06:00 [pubmed] PHST- 2020/09/23 06:00 [medline] PHST- 2019/08/16 06:00 [entrez] AID - 000502099 [pii] AID - 10.1159/000502099 [doi] PST - ppublish SO - Respiration. 2019;98(4):347-356. doi: 10.1159/000502099. Epub 2019 Aug 15.