Cargando…

Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury

Sepsis is a crucial public health problem with a high mortality rate caused by a dysregulated host immune response to infection. Vascular endothelial cell injury is an important hallmark of sepsis, which leads to multiple organ failure and death. Early biomarkers to diagnose sepsis may provide early...

Descripción completa

Detalles Bibliográficos
Autores principales: Songjang, Worawat, Paiyabhroma, Nitchawat, Jumroon, Noppadon, Jiraviriyakul, Arunya, Nernpermpisooth, Nitirut, Seenak, Porrnthanate, Kumphune, Sarawut, Thaisakun, Siriwan, Phaonakrop, Narumon, Roytrakul, Sittiruk, Pankhong, Panyupa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669054/
https://www.ncbi.nlm.nih.gov/pubmed/38002065
http://dx.doi.org/10.3390/biomedicines11113065
_version_ 1785149201973248000
author Songjang, Worawat
Paiyabhroma, Nitchawat
Jumroon, Noppadon
Jiraviriyakul, Arunya
Nernpermpisooth, Nitirut
Seenak, Porrnthanate
Kumphune, Sarawut
Thaisakun, Siriwan
Phaonakrop, Narumon
Roytrakul, Sittiruk
Pankhong, Panyupa
author_facet Songjang, Worawat
Paiyabhroma, Nitchawat
Jumroon, Noppadon
Jiraviriyakul, Arunya
Nernpermpisooth, Nitirut
Seenak, Porrnthanate
Kumphune, Sarawut
Thaisakun, Siriwan
Phaonakrop, Narumon
Roytrakul, Sittiruk
Pankhong, Panyupa
author_sort Songjang, Worawat
collection PubMed
description Sepsis is a crucial public health problem with a high mortality rate caused by a dysregulated host immune response to infection. Vascular endothelial cell injury is an important hallmark of sepsis, which leads to multiple organ failure and death. Early biomarkers to diagnose sepsis may provide early intervention and reduce risk of death. Damage-associated molecular patterns (DAMPs) are host nuclear or cytoplasmic molecules released from cells following tissue damage. We postulated that DAMPs could potentially be a novel sepsis biomarker. We used an in vitro model to determine suitable protein–DAMPs biomarkers for early sepsis diagnosis. Low and high lipopolysaccharide (LPS) doses were used to stimulate the human umbilical vein endothelial cell line EA.hy926 for 24, 48, and 72 h. Results showed that cell viability was reduced in both dose-dependent and time-dependent manners. Cell injury was corroborated by a significant increase in lactate dehydrogenase (LDH) activity within 24 h in cell-conditioned medium. Secreted protein–DAMPs in the supernatant, collected at different time points within 24 h, were characterized using shotgun proteomics LC-MS/MS analysis. Results showed that there were 2233 proteins. Among these, 181 proteins from the LPS-stimulated EA.hy926 at 1, 12, and 24 h were significantly different from those of the control. Twelve proteins were up-regulated at all three time points. Furthermore, a potential interaction analysis of predominant DAMPs-related proteins using STITCH 5.0 revealed the following associations with pathways: response to stress; bacterium; and LPS (GO:0080134; 0009617; 0032496). Markedly, alpha-2-HS-glycoprotein (AHSG or fetuin-A) and lactotransferrin (LTF) potentially presented since the first hour of LPS stimulation, and were highly up-regulated at 24 h. Taken together, we reported proteomic profiling of vascular endothelial cell-specific DAMPs in response to early an in vitro LPS stimulation, suggesting that these early damage-response protein candidates could be novel early biomarkers associated with sepsis.
format Online
Article
Text
id pubmed-10669054
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-106690542023-11-15 Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury Songjang, Worawat Paiyabhroma, Nitchawat Jumroon, Noppadon Jiraviriyakul, Arunya Nernpermpisooth, Nitirut Seenak, Porrnthanate Kumphune, Sarawut Thaisakun, Siriwan Phaonakrop, Narumon Roytrakul, Sittiruk Pankhong, Panyupa Biomedicines Article Sepsis is a crucial public health problem with a high mortality rate caused by a dysregulated host immune response to infection. Vascular endothelial cell injury is an important hallmark of sepsis, which leads to multiple organ failure and death. Early biomarkers to diagnose sepsis may provide early intervention and reduce risk of death. Damage-associated molecular patterns (DAMPs) are host nuclear or cytoplasmic molecules released from cells following tissue damage. We postulated that DAMPs could potentially be a novel sepsis biomarker. We used an in vitro model to determine suitable protein–DAMPs biomarkers for early sepsis diagnosis. Low and high lipopolysaccharide (LPS) doses were used to stimulate the human umbilical vein endothelial cell line EA.hy926 for 24, 48, and 72 h. Results showed that cell viability was reduced in both dose-dependent and time-dependent manners. Cell injury was corroborated by a significant increase in lactate dehydrogenase (LDH) activity within 24 h in cell-conditioned medium. Secreted protein–DAMPs in the supernatant, collected at different time points within 24 h, were characterized using shotgun proteomics LC-MS/MS analysis. Results showed that there were 2233 proteins. Among these, 181 proteins from the LPS-stimulated EA.hy926 at 1, 12, and 24 h were significantly different from those of the control. Twelve proteins were up-regulated at all three time points. Furthermore, a potential interaction analysis of predominant DAMPs-related proteins using STITCH 5.0 revealed the following associations with pathways: response to stress; bacterium; and LPS (GO:0080134; 0009617; 0032496). Markedly, alpha-2-HS-glycoprotein (AHSG or fetuin-A) and lactotransferrin (LTF) potentially presented since the first hour of LPS stimulation, and were highly up-regulated at 24 h. Taken together, we reported proteomic profiling of vascular endothelial cell-specific DAMPs in response to early an in vitro LPS stimulation, suggesting that these early damage-response protein candidates could be novel early biomarkers associated with sepsis. MDPI 2023-11-15 /pmc/articles/PMC10669054/ /pubmed/38002065 http://dx.doi.org/10.3390/biomedicines11113065 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Songjang, Worawat
Paiyabhroma, Nitchawat
Jumroon, Noppadon
Jiraviriyakul, Arunya
Nernpermpisooth, Nitirut
Seenak, Porrnthanate
Kumphune, Sarawut
Thaisakun, Siriwan
Phaonakrop, Narumon
Roytrakul, Sittiruk
Pankhong, Panyupa
Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title_full Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title_fullStr Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title_full_unstemmed Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title_short Proteomic Profiling of Early Secreted Proteins in Response to Lipopolysaccharide-Induced Vascular Endothelial Cell EA.hy926 Injury
title_sort proteomic profiling of early secreted proteins in response to lipopolysaccharide-induced vascular endothelial cell ea.hy926 injury
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669054/
https://www.ncbi.nlm.nih.gov/pubmed/38002065
http://dx.doi.org/10.3390/biomedicines11113065
work_keys_str_mv AT songjangworawat proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT paiyabhromanitchawat proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT jumroonnoppadon proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT jiraviriyakularunya proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT nernpermpisoothnitirut proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT seenakporrnthanate proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT kumphunesarawut proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT thaisakunsiriwan proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT phaonakropnarumon proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT roytrakulsittiruk proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury
AT pankhongpanyupa proteomicprofilingofearlysecretedproteinsinresponsetolipopolysaccharideinducedvascularendothelialcelleahy926injury