Matrilin-3 Induction of IL-1 receptor antagonist Is required for up-regulating collagen II and aggrecan and down-regulating ADAMTS-5 gene expressionReport as inadecuate




Matrilin-3 Induction of IL-1 receptor antagonist Is required for up-regulating collagen II and aggrecan and down-regulating ADAMTS-5 gene expression - Download this document for free, or read online. Document in PDF available to download.

Arthritis Research and Therapy

, 14:R197

First Online: 11 September 2012Received: 30 March 2012Revised: 28 July 2012Accepted: 21 August 2012

Abstract

IntroductionDeletion or mutation of the gene encoding the cartilage extracellular matrix ECM protein matrilin-3 MATN3 results in the early onset of osteoarthritis OA, suggesting chondroprotective properties of MATN3. To understand the mechanisms underlying these properties, we determined the effects of MATN3 protein on the expression of several key anabolic and catabolic genes involved in chondrocyte homeostasis, and the dependence of such regulation on the anti-inflammatory cytokine: IL-1 receptor antagonist IL-1Ra.

MethodsThe effects of recombinant human rh MATN3 protein were examined in C28-I2 immortalized human chondrocytes, primary human chondrocytes PHCs, and primary mouse chondrocytes PMCs. Messenger RNA levels of IL-1Ra, COL2A1, ACAN, MMP-13, and ADAMTS-4 and -5 were determined using real-time RT-PCR. Knocking down IL-1Ra was achieved by siRNA gene silencing. IL-1Ra protein levels were quantified by ELISA and the Bio-Plex Suspension Array System. COL2A1 protein level was quantified using Western blot analysis. Statistic analysis was done using the two-tailed t-test or one-way ANOVA.

ResultsrhMATN3 protein induced gene expression of IL-1Ra in C28-I2 cells, PHCs, and PMCs in a dose- and time-dependent manner. Treatment of C28-I2 cells and PHCs with MATN3 protein stimulated gene expression of COL2A1 and ACAN. Conversely, mRNA levels of COL2A1 and ACAN were decreased in MATN3 KO mice. MATN3 protein treatment inhibited IL-1β-induced MMP-13, ADAMTS-4 and ADAMTS-5 in C28-I2 cells and PHCs. Knocking down IL-1Ra abolished the MATN3-mediated stimulation of COL2A1 and ACAN and inhibition of ADAMTS-5, but had no effect on MATN3 inhibition of MMP-13 mRNA.

ConclusionOur findings point to a novel regulatory role of MATN3 in cartilage homeostasis due to its capacity to induce IL-1Ra, to upregulate gene expression of the major cartilage matrix components, and to downregulate the expression of OA-associated matrix-degrading proteinases in chondrocytes. The chondroprotective properties of endogenous MATN3 depend partly on its induction of IL-1Ra. Our findings raise a possibility to use rhMATN3 protein for anti-inflammatory and chondroprotective therapy.

KeywordsMatrilin-3 interleukin-1 IL-1Ra collagen 2 aggrecan MMP-13 ADAMTS-4 ADAMTS-5 chondrocytes osteoarthritis AbbreviationsACANaggrecan

ANOVAone-way analysis of variance

COL2A1type II collagen, DMEM: Dubecco-s modified Eagle-s medium

ECMextracellular matrix, EGF: epidermal growth factor

ELISAenzyme-linked immunosorbent assay

FBSfetal bovine serum

HBSSHank-s Balanced Salt Solution

IL-1βinterleukin-1 beta

IL-1Rainterleukin-1 receptor antagonist

KOknockout

MATN3matrilin-3

MEDmultiple epiphyseal dysplasia

MMPmatrix metalloproteinase

NOnitric oxide

OAosteoarthritis

PBSphosphate-buffered saline

PHCprimary human chondrocyte

PMCprimary mouse chondrocyte

RT-qPCRreal time quantitative polymerase chain reaction

rhrecombinant human

SEMDspondylo-epi-metaphyseal dysplasia

sIL-1Rasoluble interleukin-1 receptor antagonist

siRNAsmall interfering ribonucleic acid

vWFAVon Willebrand Factor A.

Electronic supplementary materialThe online version of this article doi:10.1186-ar4033 contains supplementary material, which is available to authorized users.

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Author: Chathuraka T Jayasuriya - Mary B Goldring - Richard Terek - Qian Chen

Source: https://link.springer.com/







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