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Effect of tryptase inhibition on joint inflammation: a pharmacological and lentivirus-mediated gene transfer study.

ABSTRACT: Increasing evidences indicate that an unbalance between tryptases and their endogenous inhibitors, leading to an increased proteolytic activity, is implicated in the pathophysiology of rheumatoid arthritis. The aim of the present study was to evaluate the impact of tryptase inhibition on experimental arthritis.Analysis of gene expression and regulation in the mouse knee joint was performed by RT-qPCR and in situ hybridization. Arthritis was induced in male C57BL/6 mice with mBSA/IL-1?. Tryptase was inhibited by two approaches: a lentivirus-mediated heterologous expression of the human endogenous tryptase inhibitor, sperm-associated antigen 11B isoform C (hSPAG11B/C), or a chronic treatment with the synthetic tryptase inhibitor APC366. Several inflammatory parameters were evaluated, such as oedema formation, histopathology, production of IL-1?, -6, -17A and CXCL1/KC, myeloperoxidase and tryptase-like activities.Spag11c was constitutively expressed in chondrocytes and cells from the synovial membrane in mice, but its expression did not change 7 days after the induction of arthritis, while tryptase expression and activity were upregulated. The intra-articular transduction of animals with the lentivirus phSPAG11B/C or the treatment with APC366 inhibited the increase of tryptase-like activity, the late phase of oedema formation, the production of IL-6 and CXCL1/KC. In contrast, neutrophil infiltration, degeneration of hyaline cartilage and erosion of subchondral bone were not affected.Tryptase inhibition was effective in inhibiting some inflammatory parameters associated to mBSA/IL-1?-induced arthritis, notably late phase oedema formation and IL-6 production, but not neutrophil infiltration and joint degeneration. These results suggest that the therapeutic application of tryptase inhibitors to rheumatoid arthritis would be restrained to palliative care, but not as disease-modifying drugs. Finally, this study highlighted lentivirus-based gene delivery as an instrumental tool to study the relevance of target genes in synovial joint physiology and disease.

SUBMITTER: Denadai-Souza A

PROVIDER: S-EPMC5461776 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Effect of tryptase inhibition on joint inflammation: a pharmacological and lentivirus-mediated gene transfer study.

Denadai-Souza Alexandre A Ribeiro Camilla Moreira CM Rolland Corinne C Thouard Anne A Deraison Céline C Scavone Cristoforo C Gonzalez-Dunia Daniel D Vergnolle Nathalie N Avellar Maria Christina Werneck MCW

Arthritis research & therapy 20170606 1

<h4>Background</h4>Increasing evidences indicate that an unbalance between tryptases and their endogenous inhibitors, leading to an increased proteolytic activity, is implicated in the pathophysiology of rheumatoid arthritis. The aim of the present study was to evaluate the impact of tryptase inhibition on experimental arthritis.<h4>Methods</h4>Analysis of gene expression and regulation in the mouse knee joint was performed by RT-qPCR and in situ hybridization. Arthritis was induced in male C57B ...[more]

PMID: 28587618

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Project description:Fibroblast growth factor 18 (FGF-18) is a well-characterized anabolic growth factor involved in cartilage homeostasis. However, the effect of FGF-18 on intervertebral disc (IVD) degeneration has not been investigated. The present study aimed to investigate the role of FGF-18 in the process of rabbit IVD degeneration. In vitro, primary nucleus pulposus cells (NPs) were cultured and transfected with a lentivirus. Tert-butyl hydroperoxide (TBHP) was used to induce apoptosis in NPs on the second passage, while overexpression of FGF-18 in NPs attenuated TBHP-induced apoptosis. A rabbit annular puncture model was generated to induce IVD degeneration in vivo. The discs were injected with an FGF-18-overexpression lentivirus or a negative control lentivirus. In the sham group, the discs were exposed and not punctured. Disc degeneration was evaluated using H&E staining and a histological grading system. Reverse transcription-quantitative PCR was used to detect the expression of the extracellular matrix-degrading enzymes matrix metalloproteinase-3 (MMP-3) and A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5). Nucleus pulposus apoptosis was detected via western blotting, immunohistochemical methods and TUNEL staining. Histologic examination showed that disc degeneration was attenuated after FGF-18 overexpression treatment. At 8 weeks after surgery, the expression of MMP-3 and ADAMTS-5 in the annular puncture groups was higher compared with in the sham group. FGF-18 treatment inhibited the expression of MMP-3 and ADAMTS-5 at the mRNA level. Western blot assays indicated that the expression level of Bax was significantly reduced in the FGF-18 groups, and that the expression level of Bcl-2 was significantly increased compared with those in the control group. Moreover, immunohistochemical analysis indicated that the FGF-18 group exhibited a lower percentage of cleaved caspase 3-positive NPs. Quantification of the TUNEL staining demonstrated that the FGF-18 group had fewer apoptotic NPs than the control group. These findings indicated that FGF-18 could delay IVD degeneration by inhibiting the apoptosis of NPs and the expression of matrix-degrading enzymes.

Dataset Information

Effect of tryptase inhibition on joint inflammation: a pharmacological and lentivirus-mediated gene transfer study.

Publications

Effect of tryptase inhibition on joint inflammation: a pharmacological and lentivirus-mediated gene transfer study.

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