Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model

BackgroundPolytrauma and hemorrhagic shock can lead to direct and indirect liver damage involving intricate pathophysiologic mechanisms. While hepatic function has been frequently highlighted, there is minimal research on how the receptor activator of the NF-κB (RANK)/RANK ligand (RANKL)/osteoproteg...

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Main Authors: Yang Li, Klemens Horst, Johannes Greven, Ümit Mert, Ludmila Lupu, Annette Palmer, Lena Doerfer, Qun Zhao, Xing Zhang, Rebecca Halbgebauer, Anita Ignatius, Ingo Marzi, Martijn van Griensven, Elizabeth Balmayor, Frank Hildebrand, Tom Eirik Mollnes, Markus Huber-Lang
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Language:English
Published: Frontiers Media S.A. 2024-11-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2024.1434274/full
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author Yang Li
Klemens Horst
Johannes Greven
Ümit Mert
Ludmila Lupu
Annette Palmer
Lena Doerfer
Qun Zhao
Xing Zhang
Rebecca Halbgebauer
Anita Ignatius
Ingo Marzi
Martijn van Griensven
Elizabeth Balmayor
Frank Hildebrand
Tom Eirik Mollnes
Tom Eirik Mollnes
Markus Huber-Lang
author_facet Yang Li
Klemens Horst
Johannes Greven
Ümit Mert
Ludmila Lupu
Annette Palmer
Lena Doerfer
Qun Zhao
Xing Zhang
Rebecca Halbgebauer
Anita Ignatius
Ingo Marzi
Martijn van Griensven
Elizabeth Balmayor
Frank Hildebrand
Tom Eirik Mollnes
Tom Eirik Mollnes
Markus Huber-Lang
author_sort Yang Li
collection DOAJ
description BackgroundPolytrauma and hemorrhagic shock can lead to direct and indirect liver damage involving intricate pathophysiologic mechanisms. While hepatic function has been frequently highlighted, there is minimal research on how the receptor activator of the NF-κB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system is regulated in the liver following trauma. Furthermore, cross-talking complement and toll-like-receptor (TLR) systems can contribute to the posttraumatic response. Therefore, we investigated the hepatic consequences of polytrauma focusing on the RANK-RANKL-OPG axis, and evaluated the effects of a dual blockade of complement factor C5 and TLR-cofactor CD14 on hepatic features.MethodsThe established pig model of polytrauma (PT) and hemorrhagic shock included pulmonary contusion, hepatic dissection, and bilateral femur fractures, surgically addressed either by external fixation (Fix ex) or intramedullary nailing (Nail). Four groups were investigated: 1) sham animals; 2) PT treated by Fix ex (Fix ex); 3) PT by Nail (Nail); or 4) PT by Nail plus combined C5/CD14 inhibition (Nail+Therapy). Serum samples were obtained between 0 - 72 h, and liver samples at 72 h after PT. Liver tissues were histologically scored and subjected to RT-qPCR-analyses, immunohistochemistry and ELISAs to evaluate the posttraumatic hepatic response with a focus on the RANK-RANKL-OPG system.ResultsFollowing PT, the liver injury score of the Nail+Therapy group was significantly lower than in the Fix ex or Nail group without immunomodulation (p<0.05). Similarly, the degree of necrosis, lobular stasis, and inflammation were significantly reduced when treated with C5/CD14-inhibitors. Compared to the Nail group, AST serum concentrations were significantly decreased in the Nail+Therapy group after 72 h (p<0.05). PCR analyses indicated that RANK, RANKL, and OPG levels in the liver were increased after PT in the Nail group compared to lower levels in the Nail+Therapy group. Furthermore, liver tissue analyses revealed increased RANK protein levels and cellular immunostaining for RANK in the Nail group, both of which were significantly reduced in the case of C5/CD14-inhibition (p<0.05).ConclusionFollowing experimental PT, dual inhibition of C5/CD14 resulted in altered, mainly reduced hepatic synthesis of proteins relevant to bone repair. However, a comprehensive investigation of the subsequent effects on the liver-bone axis are needed.
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spelling doaj-art-0dd4a07f570e42239e70c8a2eeb84d2e2024-11-21T06:23:59ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-11-011510.3389/fimmu.2024.14342741434274Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma modelYang Li0Klemens Horst1Johannes Greven2Ümit Mert3Ludmila Lupu4Annette Palmer5Lena Doerfer6Qun Zhao7Xing Zhang8Rebecca Halbgebauer9Anita Ignatius10Ingo Marzi11Martijn van Griensven12Elizabeth Balmayor13Frank Hildebrand14Tom Eirik Mollnes15Tom Eirik Mollnes16Markus Huber-Lang17Institute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyInstitute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyInstitute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyInstitute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyInstitute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyInstitute of Orthopaedic Research and Biomechanics, Ulm University Medical Center, Ulm, GermanyDepartment of Trauma, Hand, and Reconstructive Surgery, University Hospital Frankfurt, Goethe University, Frankfurt/Main, GermanyThe Department of Cell Biology-Inspired Tissue Engineering (cBITE), MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Maastricht, NetherlandsDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyDepartment of Orthopaedics, Trauma and Reconstructive Surgery, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, GermanyDepartment of Immunology, Oslo University Hospital, University of Oslo, Oslo, NorwayResearch Laboratory, Nordland Hospital Bodø, Bodø, NorwayInstitute of Clinical and Experimental Trauma Immunology, Ulm University Medical Center, Ulm, GermanyBackgroundPolytrauma and hemorrhagic shock can lead to direct and indirect liver damage involving intricate pathophysiologic mechanisms. While hepatic function has been frequently highlighted, there is minimal research on how the receptor activator of the NF-κB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system is regulated in the liver following trauma. Furthermore, cross-talking complement and toll-like-receptor (TLR) systems can contribute to the posttraumatic response. Therefore, we investigated the hepatic consequences of polytrauma focusing on the RANK-RANKL-OPG axis, and evaluated the effects of a dual blockade of complement factor C5 and TLR-cofactor CD14 on hepatic features.MethodsThe established pig model of polytrauma (PT) and hemorrhagic shock included pulmonary contusion, hepatic dissection, and bilateral femur fractures, surgically addressed either by external fixation (Fix ex) or intramedullary nailing (Nail). Four groups were investigated: 1) sham animals; 2) PT treated by Fix ex (Fix ex); 3) PT by Nail (Nail); or 4) PT by Nail plus combined C5/CD14 inhibition (Nail+Therapy). Serum samples were obtained between 0 - 72 h, and liver samples at 72 h after PT. Liver tissues were histologically scored and subjected to RT-qPCR-analyses, immunohistochemistry and ELISAs to evaluate the posttraumatic hepatic response with a focus on the RANK-RANKL-OPG system.ResultsFollowing PT, the liver injury score of the Nail+Therapy group was significantly lower than in the Fix ex or Nail group without immunomodulation (p<0.05). Similarly, the degree of necrosis, lobular stasis, and inflammation were significantly reduced when treated with C5/CD14-inhibitors. Compared to the Nail group, AST serum concentrations were significantly decreased in the Nail+Therapy group after 72 h (p<0.05). PCR analyses indicated that RANK, RANKL, and OPG levels in the liver were increased after PT in the Nail group compared to lower levels in the Nail+Therapy group. Furthermore, liver tissue analyses revealed increased RANK protein levels and cellular immunostaining for RANK in the Nail group, both of which were significantly reduced in the case of C5/CD14-inhibition (p<0.05).ConclusionFollowing experimental PT, dual inhibition of C5/CD14 resulted in altered, mainly reduced hepatic synthesis of proteins relevant to bone repair. However, a comprehensive investigation of the subsequent effects on the liver-bone axis are needed.https://www.frontiersin.org/articles/10.3389/fimmu.2024.1434274/fullpolytraumaRANK-RANKL-OPG pathwayimmunomodulationcomplementCD14
spellingShingle Yang Li
Klemens Horst
Johannes Greven
Ümit Mert
Ludmila Lupu
Annette Palmer
Lena Doerfer
Qun Zhao
Xing Zhang
Rebecca Halbgebauer
Anita Ignatius
Ingo Marzi
Martijn van Griensven
Elizabeth Balmayor
Frank Hildebrand
Tom Eirik Mollnes
Tom Eirik Mollnes
Markus Huber-Lang
Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
Frontiers in Immunology
polytrauma
RANK-RANKL-OPG pathway
immunomodulation
complement
CD14
title Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
title_full Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
title_fullStr Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
title_full_unstemmed Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
title_short Modulation of the hepatic RANK-RANKL-OPG axis by combined C5 and CD14 inhibition in a long-term polytrauma model
title_sort modulation of the hepatic rank rankl opg axis by combined c5 and cd14 inhibition in a long term polytrauma model
topic polytrauma
RANK-RANKL-OPG pathway
immunomodulation
complement
CD14
url https://www.frontiersin.org/articles/10.3389/fimmu.2024.1434274/full
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