Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways
Objective: Intervertebral disc (IVD) degeneration and disc herniation are major causes of lower back pain, which involve the presence of inflammatory mediators and tissue invasion by immune cells. Intercellular adhesion molecule 1 (ICAM1, also termed CD54) is an adhesion molecule that mediates cell-...
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doaj-5f18eae9567e467b98fca20e7dd0b4a12020-11-24T22:05:31ZengMDPI AGInternational Journal of Molecular Sciences1422-00672018-12-012017710.3390/ijms20010077ijms20010077Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling PathwaysBor-Ren Huang0Da-Tian Bau1Tzu-Sheng Chen2I-Chen Chuang3Cheng-Fang Tsai4Pei-Chun Chang5Horng-Chaung Hsu6Dah-Yuu Lu7Graduate Institute of Clinical Medical Science, China Medical University, Taichung 40402, TaiwanGraduate Institute of Clinical Medical Science, China Medical University, Taichung 40402, TaiwanDepartment of Pathology, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung 42743, TaiwanDepartment of Pharmacology, School of Medicine, China Medical University, Taichung 40402, TaiwanDepartment of Biotechnology, Asia University, Taichung 41354, TaiwanDepartment of Bioinformatics, Asia University, Taichung 41354, TaiwanDepartment of Orthopedic Surgery, China Medical University Hospital, Taichung 40402, TaiwanDepartment of Pharmacology, School of Medicine, China Medical University, Taichung 40402, TaiwanObjective: Intervertebral disc (IVD) degeneration and disc herniation are major causes of lower back pain, which involve the presence of inflammatory mediators and tissue invasion by immune cells. Intercellular adhesion molecule 1 (ICAM1, also termed CD54) is an adhesion molecule that mediates cell-cell interactions, particularly between immune cells and target tissue. The aim of this study was to examine the intracellular signaling pathways involved in inflammatory stimuli-induced ICAM1 expression in human anulus fibrosus (AF) cells. Methods: Quantitative reverse transcription-polymerase chain reaction (qPCR), western blotting, and flow cytometry were performed to dissect the roles of different signaling pathways in inflammatory stimuli-mediated ICAM1 expression. Results: Using qPCR and western blot analyses, a significant increase in ICAM1 expression was observed in AF cells after stimulation of lipopolysaccharide (LPS) plus interferon-gamma (IFNγ) in a time-dependent manner. Flow cytometry revealed ICAM1 upregulation on the surface of AF cells. Importantly, LPS plus IFNγ treatment also significantly promoted Chemokine ligand (CCL)2 expression, but not CCL3. The enhanced ICAM1 expression was abolished after incubation with antibody against CCL2. In AF cells, treatment with LPS plus IFNγ activated the FAK/ERK/GSK3 signaling pathways, promoted a time-dependent increase in PKCδ phosphorylation, and promoted PKCδ translocation to the nucleus. Treatment with the pharmacological PKCδ inhibitor; rottlerin, effectively blocked the enhanced productions of ICAM1 and CCL2. Conclusions: Inflammatory stimuli in AF cells are part of a specific pathophysiology in IVD degeneration and disc herniation that modulates CCL2/ICAM1 activation through the FAK/ERK/GSK3 and PKCδ signaling pathways in AF cells.http://www.mdpi.com/1422-0067/20/1/77intervertebral discdegenerationanulus fibrosusCCL2ICAM1 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Bor-Ren Huang Da-Tian Bau Tzu-Sheng Chen I-Chen Chuang Cheng-Fang Tsai Pei-Chun Chang Horng-Chaung Hsu Dah-Yuu Lu |
spellingShingle |
Bor-Ren Huang Da-Tian Bau Tzu-Sheng Chen I-Chen Chuang Cheng-Fang Tsai Pei-Chun Chang Horng-Chaung Hsu Dah-Yuu Lu Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways International Journal of Molecular Sciences intervertebral disc degeneration anulus fibrosus CCL2 ICAM1 |
author_facet |
Bor-Ren Huang Da-Tian Bau Tzu-Sheng Chen I-Chen Chuang Cheng-Fang Tsai Pei-Chun Chang Horng-Chaung Hsu Dah-Yuu Lu |
author_sort |
Bor-Ren Huang |
title |
Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways |
title_short |
Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways |
title_full |
Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways |
title_fullStr |
Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways |
title_full_unstemmed |
Pro-Inflammatory Stimuli Influence Expression of Intercellular Adhesion Molecule 1 in Human Anulus Fibrosus Cells through FAK/ERK/GSK3 and PKCδ Signaling Pathways |
title_sort |
pro-inflammatory stimuli influence expression of intercellular adhesion molecule 1 in human anulus fibrosus cells through fak/erk/gsk3 and pkcδ signaling pathways |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2018-12-01 |
description |
Objective: Intervertebral disc (IVD) degeneration and disc herniation are major causes of lower back pain, which involve the presence of inflammatory mediators and tissue invasion by immune cells. Intercellular adhesion molecule 1 (ICAM1, also termed CD54) is an adhesion molecule that mediates cell-cell interactions, particularly between immune cells and target tissue. The aim of this study was to examine the intracellular signaling pathways involved in inflammatory stimuli-induced ICAM1 expression in human anulus fibrosus (AF) cells. Methods: Quantitative reverse transcription-polymerase chain reaction (qPCR), western blotting, and flow cytometry were performed to dissect the roles of different signaling pathways in inflammatory stimuli-mediated ICAM1 expression. Results: Using qPCR and western blot analyses, a significant increase in ICAM1 expression was observed in AF cells after stimulation of lipopolysaccharide (LPS) plus interferon-gamma (IFNγ) in a time-dependent manner. Flow cytometry revealed ICAM1 upregulation on the surface of AF cells. Importantly, LPS plus IFNγ treatment also significantly promoted Chemokine ligand (CCL)2 expression, but not CCL3. The enhanced ICAM1 expression was abolished after incubation with antibody against CCL2. In AF cells, treatment with LPS plus IFNγ activated the FAK/ERK/GSK3 signaling pathways, promoted a time-dependent increase in PKCδ phosphorylation, and promoted PKCδ translocation to the nucleus. Treatment with the pharmacological PKCδ inhibitor; rottlerin, effectively blocked the enhanced productions of ICAM1 and CCL2. Conclusions: Inflammatory stimuli in AF cells are part of a specific pathophysiology in IVD degeneration and disc herniation that modulates CCL2/ICAM1 activation through the FAK/ERK/GSK3 and PKCδ signaling pathways in AF cells. |
topic |
intervertebral disc degeneration anulus fibrosus CCL2 ICAM1 |
url |
http://www.mdpi.com/1422-0067/20/1/77 |
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