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Interleukin-35 (IL-35) inhibits proliferation and promotes apoptosis of fibroblast-like synoviocytes isolated from mice with collagen-induced arthritis

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Abstract

Rheumatoid arthritis (RA) is an inflammatory disorder of the joints that affects 0.5–1 % of adults. Excessive growth of the fibroblast-like synoviocytes (FLS) promotes hyperplasia of synovial tissues and causes its invasion into the bone and cartilage, which eventually causes deformity and dysfunction of affected joints. Interleukin 35 (IL-35) was shown to suppress the inflammatory responses to collagen-induced arthritis (CIA) via upregulation of T regulatory cells and suppression of T helper type 17 cells in a mouse model. To study the effects of IL-35 on the proliferation and apoptosis frequency of cultured FLS isolated from mice with CIA as well as to examine the effects of IL-35 on CIA in vivo. Thirty DBA/1 J mice, which are used as an animal model for RA, were divided randomly (ten mice per group) to a CIA group (collagen treatment), a CIA + IL-35 group (collagen and IL-35 treatments), and a control group (no treatment). Starting on the 24th day after collagen administration, IL-35 was injected intraperitoneally into mice of the CIA + IL-35 group once per day for 10 days. An arthritis index was calculated, and pathological analysis of synovial tissue was performed. FLS isolated from CIA mice were treated with various concentrations of IL-35 (12.5–100 ng/ml). The MTT assay was used to examine FLS proliferation, and apoptosis frequency of FLS was detected by flow cytometry. On day 24, the CIA mice began to exhibit arthritis symptoms, and the symptoms rapidly progressed with time. Treatment with IL-35 significantly alleviated arthritis symptoms and reduced the synovial tissue inflammation. In addition, IL-35 treatment inhibited proliferation and promoted apoptosis in cultured FLS from CIA mice in a dose-dependent manner. IL-35 could ameliorate the symptoms of arthritis in the CIA mouse model in vivo and inhibited FLS proliferation while promoting FLS apoptosis in vitro, thereby exhibited the potential in inhibiting the progression of RA.

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Abbreviations

RA:

Rheumatoid arthritis

FLS:

Fibroblast-like synoviocytes

IL-35:

Interleukin-35

Th17:

T helper type 17

CIA:

Collagen-induced arthritis

MTT:

3-(4,5-Dimethyl-thiazoyl-2-yl)-2,5-diphenyltetrazolium bromide

MMPs:

Matrix metalloproteinases

IACUC:

Institutional Animal Care and Use Committee

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

H2O2 :

Hydrogen peroxide

DAB:

3,3-Diaminobenzidine

SDS-PAGE:

Sodium dodecylsulphate-polyarcylamide gel electrophoresis

ANOVA:

One-way analysis of variance

SD:

Standard deviation

LSD:

Least significant difference

Ao-SMCs:

Human aortic smooth muscle cells

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Acknowledgments

This work was supported by Grants from the National Natural Science Foundation of China (Nos. 81172867, 81471542).

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Correspondence to Jing Lu.

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Li, Y., Wu, S., Li, Y. et al. Interleukin-35 (IL-35) inhibits proliferation and promotes apoptosis of fibroblast-like synoviocytes isolated from mice with collagen-induced arthritis. Mol Biol Rep 43, 947–956 (2016). https://doi.org/10.1007/s11033-016-4034-7

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  • DOI: https://doi.org/10.1007/s11033-016-4034-7

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