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S100A6 Activates Kupffer Cells via the p-P38 and p-JNK Pathways to Induce Inflammation, Mononuclear/macrophage Infiltration Sterile Liver Injury in Mice

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A Correction to this article was published on 09 January 2023

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Abstract

Noninfectious liver injury, including the effects of chemical material, drugs and diet, is a major cause of liver diseases worldwide. In chemical and drugs-induced liver injury, innate inflammatory responses are mediated by extracellular danger signals. The S100 protein can act as danger signals, which can promote the migration and chemotaxis of immune cells, promote the release of various inflammatory cytokines, and regulate the body’s inflammatory and immune responses. However, the role of S100A6 in inflammatory response in chemical and drugs-induced sterile liver injury remains unclear. We constructed the model of sterile liver injury induced by carbon tetrachloride (CCl4)/Paracetamol (APAP) and performed RNA sequencing (RNA-seq) on the liver tissues after injury (days 2 and 5). We analyzed inflammatory protein secretion in the liver tissue supernatant by enzyme-linked immunosorbent assay (ELISA), determined the inflammation response by bioinformatic analysis during sterile liver injury, and assessed mononuclear/macrophage infiltration by immunohistochemistry and flow cytometry. Immunohistochemistry was used to analyze the location of S100A6. We conducted inflammatory factor expression analysis and molecular mechanistic studies in Kupffer cells (KCs) induced by S100A6 using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), ELISA, and western blot in vitro experiments. We performed chemokine CCL2 expression analysis and molecular mechanism studies using the same method. We used a Transwell assay to show the infiltration of mononuclear/macrophage. We here observed that aggravated inflammatory response was shown in CCl4 and APAP-administrated mice, as evidenced by enhanced production of inflammatory cytokines (TNF-α, IL-1β), and elevated mononuclear/macrophage infiltration and activation of immunity. The expression of S100A6 was significantly increased on day 2 after sterile liver injury, which is primarily produced by injured liver cells. Mechanistic studies established that S100A6 activates Kupffer cells (KCs) via the p-P38, p-JNK and P65 pathways to induce inflammation in vitro. Furthermore, TNF-α can stimulate liver cells via the p-P38 and p-JNK pathways to produce CCL2 and promote the infiltration of mononuclear/macrophage. In summary, we showed that S100A6 plays an important role in regulating inflammation, thus influencing sterile liver injury. Our findings provide novel evidence that S100A6 can as a danger signal that contributes to pro-inflammatory activation through p-P38 and p-JNK pathways in CCl4 and APAP-induced sterile liver injury in mice. In addition, the inflammatory factor TNF-α induces a large amount of CCL2 production in normal liver cells surrounding the injured area through a paracrine action, which is chemotactic for blood mononuclear/macrophage infiltration.

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AVAILABILITY OF DATA AND MATERIALS

The dataset supporting the conclusions of this article are included within the article.

Change history

Abbreviations

ALT:

Alanine transaminase

AST:

Aspartate transaminase

CCL2:

Chemokine (C–C motif) ligand 2

ELISA:

Enzyme-linked immunosorbent assay

ERK:

Extracellular regulated protein kinases

GO:

Gene Ontology

HE:

Hematoxylin and eosin

JNK:

C-Jun N-terminal kinase

KEGG:

Kyoto Encyclopedia of Genes and Genomes

IL-1β:

Interleukin-1β

IL-6:

Interleukin-6

STAT:

Signal transducer and activator of transcription

TNF-α:

Tumoral necrosis factor-α

TLR4:

Toll-like receptors

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ACKNOWLEDGEMENTS

We thank editage. Co., Ltd., which was used for language modifications.

Funding

This study was supported by grants from Inner Mongolia Autonomous Region Key Technology Tackle Project (2021GG0098) and Inner Mongolia Autonomous Region Cancer Biotherapy Collaborative Innovation Cultivation Center.

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Author notes

  1. He Tong and Li Wang contributed equally to this work.

Authors and Affiliations

  1. School of Life Science, Inner Mongolia University, Yu Quan District, Xin Lin Guo Le Nan Road 49, Hohhot, 010020, Inner Mongolia, China

    He Tong, Kefan Zhang, Jing Shi, Yongshuai Wu & Changshan Wang

  2. School of Basic Medicine, Inner Mongolia Medical University, Hohhot, 010110, Inner Mongolia, China

    Li Wang & Yulong Bao

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Contributions

H.T. and L.W. contributed to the conception and design of the study, acquired the majority of the data, and drafted the manuscript. K.Z. contributed to the conception and design of the study and acquired some of the data. J.S. and Y.S. contributed to the design of the study and interpretation of the data. C.W. and Y.B. contributed to the design of the study, interpretation of data, and substantively revised the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Yulong Bao or Changshan Wang.

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Ethical Approval and Consent to Participate

This study was conducted in accordance with the guidelines of the Declaration of Helsinki. All animal experimental protocols applied in this study were conducted in accordance with the standards of the Ethics Committee of Inner Mongolia Medical University (YKD202001030).

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The authors declare no competing interests.

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The original online version of this article was revised: The corresponding authors of this article should be Yulong Bao, yulongbao0471@163.com, and Changshan Wang, changshanwang@imu.edu.cn.

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Tong, H., Wang, L., Zhang, K. et al. S100A6 Activates Kupffer Cells via the p-P38 and p-JNK Pathways to Induce Inflammation, Mononuclear/macrophage Infiltration Sterile Liver Injury in Mice. Inflammation 46, 534–554 (2023). https://doi.org/10.1007/s10753-022-01750-w

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