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Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2
SIMPLE SUMMARY: In this study, the yak CXCL14 protein was successfully obtained through prokaryotic expression. In vitro experiments proved that the CXCL14 protein can not only significantly increase the activity of spleen macrophages and promote the expression of pro-inflammatory cell-related facto...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10571970/ https://www.ncbi.nlm.nih.gov/pubmed/37835641 http://dx.doi.org/10.3390/ani13193036 |
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author | Li, Biao Li, Juan Wang, Li Wei, Yong Luo, Xiaolin Guan, Jiuqiang Zhang, Xiangfei |
author_facet | Li, Biao Li, Juan Wang, Li Wei, Yong Luo, Xiaolin Guan, Jiuqiang Zhang, Xiangfei |
author_sort | Li, Biao |
collection | PubMed |
description | SIMPLE SUMMARY: In this study, the yak CXCL14 protein was successfully obtained through prokaryotic expression. In vitro experiments proved that the CXCL14 protein can not only significantly increase the activity of spleen macrophages and promote the expression of pro-inflammatory cell-related factors but also inhibit the activity, clone formation and migration of HepG2. This study aims to further understand the function of the CXCL14 protein in yaks. ABSTRACT: CXCL14 (C-X-C motif chemokine ligand 14) is an important chemokine involved in infection and immunity and plays an important role in a variety of immune-related diseases. The 446 bp cDNA sequence of the CXCL14 gene in yaks was obtained. Additionally, the prokaryotic expression vector of the CXCL14 protein with a molecular weight of 27 kDa was successfully constructed and expressed. The proliferation activities and migration abilities of spleen macrophages were significantly inhibited after treatment with the CXCL14 protein at different concentrations (1, 10 and 20 μg/mL) (p < 0.05). Furthermore, the expressions of pro-inflammatory cytokines interleukin 1 beta (IL-1β), interleukin 6 (IL6), interleukin 8 (IL8) and interferon-α (TNF-α) were significantly increased (p < 0.05), but the expression of anti-inflammatory factor interleukin 10 (IL10) was significantly decreased (p < 0.05). The contents of inflammatory factors in the supernatant of cells were detected using ELISA, and it was also found that the contents of TNF-α, IL6 and cytochrome c oxidase subunit II (COX2) were significantly increased under different CXCL14 protein concentrations (p < 0.05). Finally, the exogenous addition of CXCL14 inhibited the activity, clonal formation and migration of hepatoma cells (HepG2). Additionally, after HepG2 cells were treated with 20 μg/mL CXCL14 protein for 12 h, 24 h and 36 h, the expression levels of BCL2 homologous antagonist/killer (BAK) and the BCL2-associated X apoptosis regulator (BAX) were increased to varying degrees, while the expression levels of hypoxia-inducible factor 1 subunit alpha (HIF1A), the mechanistic target of rapamycin kinase (mTOR) and cyclin-dependent kinase 1 (CDK1) genes decreased compared to the control group. In conclusion, the CXCL14 protein can inhibit the proliferation and migration of HepG2 cells by inducing the expression of macrophage pro-inflammatory factors and activating apoptosis-related genes to exert innate immunity. These results are helpful to further study the function of the CXCL14 protein and provide research data for the innate immune mechanism of yaks under harsh plateau environments. |
format | Online Article Text |
id | pubmed-10571970 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-105719702023-10-14 Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 Li, Biao Li, Juan Wang, Li Wei, Yong Luo, Xiaolin Guan, Jiuqiang Zhang, Xiangfei Animals (Basel) Article SIMPLE SUMMARY: In this study, the yak CXCL14 protein was successfully obtained through prokaryotic expression. In vitro experiments proved that the CXCL14 protein can not only significantly increase the activity of spleen macrophages and promote the expression of pro-inflammatory cell-related factors but also inhibit the activity, clone formation and migration of HepG2. This study aims to further understand the function of the CXCL14 protein in yaks. ABSTRACT: CXCL14 (C-X-C motif chemokine ligand 14) is an important chemokine involved in infection and immunity and plays an important role in a variety of immune-related diseases. The 446 bp cDNA sequence of the CXCL14 gene in yaks was obtained. Additionally, the prokaryotic expression vector of the CXCL14 protein with a molecular weight of 27 kDa was successfully constructed and expressed. The proliferation activities and migration abilities of spleen macrophages were significantly inhibited after treatment with the CXCL14 protein at different concentrations (1, 10 and 20 μg/mL) (p < 0.05). Furthermore, the expressions of pro-inflammatory cytokines interleukin 1 beta (IL-1β), interleukin 6 (IL6), interleukin 8 (IL8) and interferon-α (TNF-α) were significantly increased (p < 0.05), but the expression of anti-inflammatory factor interleukin 10 (IL10) was significantly decreased (p < 0.05). The contents of inflammatory factors in the supernatant of cells were detected using ELISA, and it was also found that the contents of TNF-α, IL6 and cytochrome c oxidase subunit II (COX2) were significantly increased under different CXCL14 protein concentrations (p < 0.05). Finally, the exogenous addition of CXCL14 inhibited the activity, clonal formation and migration of hepatoma cells (HepG2). Additionally, after HepG2 cells were treated with 20 μg/mL CXCL14 protein for 12 h, 24 h and 36 h, the expression levels of BCL2 homologous antagonist/killer (BAK) and the BCL2-associated X apoptosis regulator (BAX) were increased to varying degrees, while the expression levels of hypoxia-inducible factor 1 subunit alpha (HIF1A), the mechanistic target of rapamycin kinase (mTOR) and cyclin-dependent kinase 1 (CDK1) genes decreased compared to the control group. In conclusion, the CXCL14 protein can inhibit the proliferation and migration of HepG2 cells by inducing the expression of macrophage pro-inflammatory factors and activating apoptosis-related genes to exert innate immunity. These results are helpful to further study the function of the CXCL14 protein and provide research data for the innate immune mechanism of yaks under harsh plateau environments. MDPI 2023-09-27 /pmc/articles/PMC10571970/ /pubmed/37835641 http://dx.doi.org/10.3390/ani13193036 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 Li, Biao Li, Juan Wang, Li Wei, Yong Luo, Xiaolin Guan, Jiuqiang Zhang, Xiangfei Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title | Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title_full | Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title_fullStr | Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title_full_unstemmed | Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title_short | Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2 |
title_sort | yak-derived cxcl14 activates the pro-inflammatory response of macrophages and inhibits the proliferation and migration of hepg2 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10571970/ https://www.ncbi.nlm.nih.gov/pubmed/37835641 http://dx.doi.org/10.3390/ani13193036 |
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