LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract
Jun Luo,1,* Li Li,2,* Die Hu,2 Xian Zhang1 1Department of Laboratory Medicine, Chengdu Second People’s Hospital, Chengdu 610000, Sichuan, People’s Republic of China; 2Department of Respiratory and Critical Care Medicine, Dujiangyan People’s Hospital, Dujiangyan 611830,...
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doaj-30411bb898c94b8b800330774566d8092020-11-25T03:43:50ZengDove Medical PressInternational Journal of COPD1178-20052020-08-01Volume 152049206056661LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke ExtractLuo JLi LHu DZhang XJun Luo,1,* Li Li,2,* Die Hu,2 Xian Zhang1 1Department of Laboratory Medicine, Chengdu Second People’s Hospital, Chengdu 610000, Sichuan, People’s Republic of China; 2Department of Respiratory and Critical Care Medicine, Dujiangyan People’s Hospital, Dujiangyan 611830, Sichuan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li LiDepartment of Respiratory and Critical Care Medicine, Dujiangyan People’s Hospital, No. 622 Baolian Road, Dujiangyan 611830, Sichuan, People’s Republic of ChinaTel +86-28-87121111Email lili20100901@163.comBackground: Long non-coding RNAs (lncRNAs) have been reported as key regulators in chronic obstructive pulmonary disease (COPD). However, the precise role of LINC00612 remains unclear.Methods: The real-time quantitative polymerase chain reaction (RT-qPCR) was used to quantify the expression levels of LINC00612, miR-31-5p, and Notch homolog 1 (Notch1) in lung tissues and cells. Under a cigarette smoke extract (CSE) stimulation condition, the apoptosis was analyzed by flow cytometry assay. Caspase-3 activity was examined with a caspase-3 activity assay kit; besides, inflammation and oxidative stress were assessed by measuring interleukin-6, tumor necrosis factor-α, glutathione/oxidized glutathione, reactive oxygen species, malondialdehyde, and superoxide dismutase activity. The interaction relationship between miR-31-5p and LINC00612 or Notch1 was predicted by bioinformatics databases, while dual-luciferase reporter, RNA immunoprecipitation, and RNA pull-down assays were performed to confirm prediction. Eventually, the related protein expression was estimated with western blot assay.Results: LINC00612 was downregulated in COPD tissues when compared with controls. Consistently, CSE inhibited LINC00612 expression in HPMECs with a dose/time-dependent method. Gain-of-function experiments indicated that the upregulation of LINC00612 could repress cell apoptosis, inflammation, and oxidative stress in HPMECs induced by CSE. In addition, miR-31-5p was negatively regulated by LINC00612 in HPMECs treated with CSE. The overexpression of miR-31-5p could abolish LINC00612-induced effects on HPMECs exposed to CSE. Importantly, LINC00612 could weaken CSE-induced cell apoptosis, inflammation, and oxidative stress in HPMECs by regulating the miR-31-5p/Notch1 signaling pathway.Conclusion: Current findings suggest that CSE-mediated cell apoptosis, inflammation, and oxidative stress in HPMECs were abolished by upregulation of LINC00612. Furthermore, the LINC00612/miR-31-5p/Notch1 axis may represent a novel regulator of apoptosis, inflammation, and oxidative stress of HPMECs, which may be a potential therapeutic target for COPD in the future.Keywords: LINC00612, miR-31-5p, Notch1, COPD, CSEhttps://www.dovepress.com/linc00612mir-31-5pnotch1-axis-regulates-apoptosis-inflammation-and-oxi-peer-reviewed-article-COPDlinc00612mir-31-5pnotch1copdcse |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Luo J Li L Hu D Zhang X |
spellingShingle |
Luo J Li L Hu D Zhang X LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract International Journal of COPD linc00612 mir-31-5p notch1 copd cse |
author_facet |
Luo J Li L Hu D Zhang X |
author_sort |
Luo J |
title |
LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract |
title_short |
LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract |
title_full |
LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract |
title_fullStr |
LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract |
title_full_unstemmed |
LINC00612/miR-31-5p/Notch1 Axis Regulates Apoptosis, Inflammation, and Oxidative Stress in Human Pulmonary Microvascular Endothelial Cells Induced by Cigarette Smoke Extract |
title_sort |
linc00612/mir-31-5p/notch1 axis regulates apoptosis, inflammation, and oxidative stress in human pulmonary microvascular endothelial cells induced by cigarette smoke extract |
publisher |
Dove Medical Press |
series |
International Journal of COPD |
issn |
1178-2005 |
publishDate |
2020-08-01 |
description |
Jun Luo,1,* Li Li,2,* Die Hu,2 Xian Zhang1 1Department of Laboratory Medicine, Chengdu Second People’s Hospital, Chengdu 610000, Sichuan, People’s Republic of China; 2Department of Respiratory and Critical Care Medicine, Dujiangyan People’s Hospital, Dujiangyan 611830, Sichuan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Li LiDepartment of Respiratory and Critical Care Medicine, Dujiangyan People’s Hospital, No. 622 Baolian Road, Dujiangyan 611830, Sichuan, People’s Republic of ChinaTel +86-28-87121111Email lili20100901@163.comBackground: Long non-coding RNAs (lncRNAs) have been reported as key regulators in chronic obstructive pulmonary disease (COPD). However, the precise role of LINC00612 remains unclear.Methods: The real-time quantitative polymerase chain reaction (RT-qPCR) was used to quantify the expression levels of LINC00612, miR-31-5p, and Notch homolog 1 (Notch1) in lung tissues and cells. Under a cigarette smoke extract (CSE) stimulation condition, the apoptosis was analyzed by flow cytometry assay. Caspase-3 activity was examined with a caspase-3 activity assay kit; besides, inflammation and oxidative stress were assessed by measuring interleukin-6, tumor necrosis factor-α, glutathione/oxidized glutathione, reactive oxygen species, malondialdehyde, and superoxide dismutase activity. The interaction relationship between miR-31-5p and LINC00612 or Notch1 was predicted by bioinformatics databases, while dual-luciferase reporter, RNA immunoprecipitation, and RNA pull-down assays were performed to confirm prediction. Eventually, the related protein expression was estimated with western blot assay.Results: LINC00612 was downregulated in COPD tissues when compared with controls. Consistently, CSE inhibited LINC00612 expression in HPMECs with a dose/time-dependent method. Gain-of-function experiments indicated that the upregulation of LINC00612 could repress cell apoptosis, inflammation, and oxidative stress in HPMECs induced by CSE. In addition, miR-31-5p was negatively regulated by LINC00612 in HPMECs treated with CSE. The overexpression of miR-31-5p could abolish LINC00612-induced effects on HPMECs exposed to CSE. Importantly, LINC00612 could weaken CSE-induced cell apoptosis, inflammation, and oxidative stress in HPMECs by regulating the miR-31-5p/Notch1 signaling pathway.Conclusion: Current findings suggest that CSE-mediated cell apoptosis, inflammation, and oxidative stress in HPMECs were abolished by upregulation of LINC00612. Furthermore, the LINC00612/miR-31-5p/Notch1 axis may represent a novel regulator of apoptosis, inflammation, and oxidative stress of HPMECs, which may be a potential therapeutic target for COPD in the future.Keywords: LINC00612, miR-31-5p, Notch1, COPD, CSE |
topic |
linc00612 mir-31-5p notch1 copd cse |
url |
https://www.dovepress.com/linc00612mir-31-5pnotch1-axis-regulates-apoptosis-inflammation-and-oxi-peer-reviewed-article-COPD |
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