Cigarette Smoke Extract Promotes Human Lung Myofibroblast Differentiation by the Induction of Endoplasmic Reticulum Stress
Cigarette Smoke Extract Promotes Human Lung Myofibroblast Differentiation by the Induction of Endoplasmic Reticulum Stress
复制标题
香烟烟雾提取物通过诱导内质网应激促进人肺肌成纤维细胞分化
DOI：
10.1159/000502099
复制
发表时间：
2019-10-01
期刊：
影响因子：
3.7
通讯作者：
Zhou, Yong
中科院分区：
文献类型：
--
作者：
Song, Min;Peng, Hong;Zhou, Yong
背景：特发性肺纤维化（IPF）是一种进行性、致命性纤维化性肺病，病因不明。持续的肌成纤维细胞分化是IPF的一个显着特征。吸烟是 IPF 的危险因素，也是预后不良的指标。吸烟会诱导内质网（ER）应激，并且研究表明，ER应激可促进肺纤维化中成纤维细胞向肌成纤维细胞的分化。在这项研究中，我们研究了香烟烟雾提取物 (CSE) 是否通过诱导 ER 应激来促进肺肌成纤维细胞分化。目的：我们的研究重点探讨吸烟与内质网应激在肺成纤维细胞向肌成纤维细胞分化过程中的关系。方法：用不同剂量的 CSE 刺激人胚胎肺成纤维细胞（MRC-5 成纤维细胞）。通过免疫荧光和蛋白质印迹分析评估α-平滑肌肌动蛋白（α-SMA）蛋白的水平。内质网应激由毒胡萝卜素 (TG) 诱导，并由 4-苯基丁酸 (4-PBA) 抑制。通过蛋白质印迹法测定葡萄糖调节蛋白 78 (GRP78)、肌醇需求酶 1 (IRE1)、X 盒结合蛋白 1 (XBP-1) 和激活转录因子 6 (ATF6) 的蛋白水平。使用 Lipofectamine RNAiMAX 试剂将 GRP78 siRNA 转染至 MRC-5 细胞中。结果：1.0%浓度的CSE显着增加MRC-5细胞中α-SMA的表达。细胞暴露于CSE后没有明显的细胞凋亡。 CSE处理48小时后蛋白水平上GRP78、IRE1、XBP-1和ATF6的表达显着增加。 TG 预处理增强 CSE 诱导的 α-SMA、GRP78 和 XBP-1 表达，而 4-PBA 预处理则抑制 CSE 诱导的 α-SMA、GRP78 和 XBP-1 表达。此外，GRP78 的敲低可阻断暴露于 CSE 的 MRC-5 细胞中的 α-SMA 表达。结论：我们的数据表明，CSE 通过诱导 ER 应激来促进肺成纤维细胞向肌成纤维细胞的分化。
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 α-smooth muscle actin (α-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 α-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 α-SMA, GRP78 and XBP-1. Furthermore, knockdown of GRP78 blocked α-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.