Denadai-Souza Alexandre, Ribeiro Camilla Moreira, Rolland Corinne, Thouard Anne, Deraison Céline, Scavone Cristoforo, Gonzalez-Dunia Daniel, Vergnolle Nathalie, Avellar Maria Christina Werneck
Department of Pharmacology, Universidade Federal de São Paulo - Escola Paulista de Medicina (UNIFESP-EPM), Rua 03 de Maio, São Paulo, 04044-020, Brazil.
IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
Arthritis Res Ther. 2017 Jun 6;19(1):124. doi: 10.1186/s13075-017-1326-9.
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-1β. 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-1β, -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-1β-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.
越来越多的证据表明，类胰蛋白酶与其内源性抑制剂之间的失衡导致蛋白水解活性增加，这与类风湿性关节炎的病理生理学有关。本研究的目的是评估类胰蛋白酶抑制对实验性关节炎的影响。
通过逆转录定量聚合酶链反应（RT-qPCR）和原位杂交对小鼠膝关节中的基因表达和调控进行分析。用甲基化牛血清白蛋白/白细胞介素-1β（mBSA/IL-1β）诱导雄性C57BL/6小鼠患关节炎。通过两种方法抑制类胰蛋白酶：慢病毒介导的人内源性类胰蛋白酶抑制剂精子相关抗原11B异构体C（hSPAG11B/C）的异源表达，或用合成类胰蛋白酶抑制剂APC366进行长期治疗。评估了几个炎症参数，如水肿形成、组织病理学、白细胞介素-1β、-6、-17A和CXC趋化因子配体1/角质形成细胞趋化因子（CXCL1/KC）的产生、髓过氧化物酶和类胰蛋白酶样活性。
Spag11c在小鼠软骨细胞和滑膜细胞中组成性表达，但在关节炎诱导7天后其表达没有变化，而类胰蛋白酶的表达和活性上调。用慢病毒phSPAG11B/C对动物进行关节内转导或用APC366治疗可抑制类胰蛋白酶样活性的增加、水肿形成的后期阶段、白细胞介素-6和CXCL1/KC的产生。相比之下，中性粒细胞浸润、透明软骨变性和软骨下骨侵蚀不受影响。
类胰蛋白酶抑制在抑制与mBSA/IL-1β诱导的关节炎相关的一些炎症参数方面有效，特别是水肿形成的后期阶段和白细胞介素-6的产生，但对中性粒细胞浸润和关节退变无效。这些结果表明，类胰蛋白酶抑制剂在类风湿性关节炎中的治疗应用将仅限于姑息治疗，而不是作为改善病情的药物。最后，本研究强调基于慢病毒的基因传递是研究靶基因在滑膜关节生理学和疾病中的相关性的一种有用工具。
