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Interleukin-35 upregulates OPG and inhibits RANKL in mice with collagen-induced arthritis and fibroblast-like synoviocytes

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Abstract

Summary

IL-35 is a novel anti-inflammatory cytokine, but the exact role of IL-35 in the progression of RA remains unclear, especially associated with osteoporosis and bone erosion. The present research has not been reported. Our purpose is to study how IL-35 affects RA bone destruction.

Introduction

This study investigated the effect of interleukin-35 (IL-35) on OPG and RANKL expression in collagen-induced arthritis (CIA) in rats and in cultured fibroblast-like synoviocytes (FLS).

Methods

Thirty DBA/1J mice were randomly assigned to three groups (n = 10 per group): the control group, the CIA group, and the CIA + IL-35 group. Collagen-induced arthritis was induced by immunization with collagen. IL-35 was intraperitoneally injected daily for 10 days, starting from the 24th day after immunization. FLS cells were isolated and cultured from CIA. The expression of IL-17, RANKL, and OPG was determined by RT-PCR and Western blot. Each experiment was repeated three times.

Results

CIA mice exhibited arthritis symptoms on day 24, followed by a rapid progression of arthritis. The expression of IL-17 and RANKL was increased and the expression of OPG was decreased in CIA mice compared with control mice. IL-35 treatment inhibited the development of arthritis in CIA mice, accompanied by a decrease in the expression of IL-17 and RANKL and an increase in the expression of OPG. Furthermore, IL-35 dose-dependently inhibited the expression of RANKL and increased the expression of OPG in cultured FLS cells.

Conclusion

IL-35 inhibits RANKL expression and increases OPG expression in CIA mice. IL-35 may be used for treating rheumatoid arthritis.

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Abbreviations

RA:

Rheumatoid arthritis

FLS:

Fibroblast-like synoviocytes

IL-35:

Interleukin-35

CIA:

Collagen-induced arthritis

RANK:

Receptor activator of nuclear factor κB

RANKL:

Receptor activator of nuclear factor κB ligand

OPG:

Osteoprotegerin

EBI:

Epstein-Barr virus-induced protein

Treg:

Regulatory T cells

Th17:

T helper type 17

CFA:

Complete Freund’s adjuvant

IFA:

Incomplete Freund’s adjuvant

FBS:

Fetal bovine serum

HE:

Hematoxylin and eosin

H-DMEM:

High glucose Dulbecco’s Modified Eagle’s medium

DAB:

3,3′-diaminobenzidine

SDS–PAGE:

Sodium dodecylsulphate-polyarcylamide gel electrophoresis

IOD:

Integratedoptical density

AA:

Accumulated area

ANOVA:

One-way analysis of variance

SD:

Standard deviation

LSD:

Least significant difference

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 81172867 and 81471542).

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Authors and Affiliations

  1. Department of Rheumatology and Immunology, The First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang, 110001, People’s Republic of China

    Y. Li, D. Li, Y. Li, S. Wu, S. Jiang, T. Lin, L. Xia, H. Shen & J. Lu

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  1. Y. Li
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Correspondence to J. Lu.

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The Institutional Animal Care and Use Committee of China Medical University approved all experimental protocols associated with this study.

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Li, Y., Li, D., Li, Y. et al. Interleukin-35 upregulates OPG and inhibits RANKL in mice with collagen-induced arthritis and fibroblast-like synoviocytes. Osteoporos Int 27, 1537–1546 (2016). https://doi.org/10.1007/s00198-015-3410-9

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  • DOI: https://doi.org/10.1007/s00198-015-3410-9

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