Interleukin-35 upregulates OPG and inhibits RANKL in mice with collagen-induced arthritis and fibroblast-like synoviocytes
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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.
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).
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.
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.
IL-35 inhibits RANKL expression and increases OPG expression in CIA mice. IL-35 may be used for treating rheumatoid arthritis.
KeywordsInterleukin-35 OPG RANKL Rheumatic arthritis
Receptor activator of nuclear factor κB
Receptor activator of nuclear factor κB ligand
Epstein-Barr virus-induced protein
Regulatory T cells
T helper type 17
Complete Freund’s adjuvant
Incomplete Freund’s adjuvant
Fetal bovine serum
Hematoxylin and eosin
High glucose Dulbecco’s Modified Eagle’s medium
Sodium dodecylsulphate-polyarcylamide gel electrophoresis
One-way analysis of variance
Least significant difference
This work was supported by grants from the National Natural Science Foundation of China (No. 81172867 and 81471542).
Compliance with ethical standards
The Institutional Animal Care and Use Committee of China Medical University approved all experimental protocols associated with this study.
Conflicts of interest
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