Granzyme A Contributes to Inflammatory Arthritis in Mice Through Stimulation of Osteoclastogenesis
Granzyme A Contributes to Inflammatory Arthritis in Mice Through Stimulation of Osteoclastogenesis
复制标题
DOI：
10.1002/art.39857
复制
发表时间：
2017-02-01
影响因子：
13.3
通讯作者：
Froelich, Christopher J.
中科院分区：
文献类型：
--
作者：
Santiago, Llipsy;Menaa, Cheikh;Froelich, Christopher J.
Objective.类风湿关节炎（RA）患者血浆和滑膜中颗粒酶A（GzmA）水平升高，表明该蛋白酶参与疾病的发病机制。GzmA通过调节促炎细胞因子的产生而导致脓毒症。本研究的目的是评估GzmA在体内RA发病机制中的作用，并研究GzmA通过肿瘤坏死因子（TNF）刺激破骨细胞生成的可能性。在野生型、GzmA缺陷型和穿孔素缺陷型小鼠中评价II型胶原诱导的炎性关节炎。体外实验采用骨髓细胞和粒细胞-巨噬细胞集落形成单位（CFU-GM）细胞，体内实验采用GzmA缺陷小鼠，检测GzmA的破骨细胞生成能力。GzmA基因缺失可减轻胶原诱导的关节炎，包括受影响小鼠的促炎细胞因子、关节损伤和骨质侵蚀的血清水平，这表明在没有GzmA的情况下破骨细胞活性降低。因此，GzmA处理的骨髓细胞产生的多核细胞满足成熟破骨细胞的标准：抗酒石酸酸性磷酸酶（TRAP）活性，整合素表达，降钙素受体表达和牙本质切片上的再吸收活性。GzmA似乎在没有辅助细胞的情况下起作用，其活性不受骨保护素的影响，表明RANKL的作用较小。并诱导TNF的表达和分泌。中和TNF或刺激来自TNF-/-小鼠的CFU-GM细胞可阻止GzmA诱导的破骨细胞生成。GzmA缺陷小鼠体内破骨细胞生成减少（降钙素受体阳性多核细胞减少，关节组织蛋白酶K、基质金属蛋白酶9和TRAP转录物减少），血清I型胶原C末端肽水平降低。GzmA通过促进破骨细胞分化参与RA的关节破坏。
Objective. Granzyme A (GzmA) levels are elevated in the plasma and synovium of patients with rheumatoid arthritis (RA), suggesting involvement of this protease in the pathogenesis of the disease. GzmA contributes to sepsis by regulating the production of proinflammatory cytokines. The purpose of this study was to evaluate the contribution of GzmA to the pathogenesis of RA in vivo and to examine the possibility that GzmA acting via tumor necrosis factor (TNF) stimulates osteoclastogenesis.Methods. Inflammatory arthritis induced by type II collagen was evaluated in wild-type, GzmA-deficient, and perforin-deficient mice. The osteoclastogenic potential of GzmA was examined in vitro using bone marrow cells and colony-forming unit-granulocyte-macrophage (CFU-GM) cells and in vivo using GzmA-deficient mice.Results. Gene deletion of GzmA attenuated collagen-induced arthritis, including serum levels of proinflammatory cytokines, joint damage, and bone erosion in affected mice, suggesting that osteoclast activity is reduced in the absence of GzmA. Accordingly, GzmA-treated bone marrow cells produced multinucleated cells that fulfilled the criteria for mature osteoclasts: tartrate-resistant acid phosphatase (TRAP) activity, integrin expression, calcitonin receptor expression, and resorptive activity on dentin slices. GzmA appeared to act without accessory cells, and its activity was not affected by osteoprotegerin, suggesting a minor contribution of RANKL. It also induced the expression and secretion of TNF. Neutralization of TNF or stimulation of CFU-GM cells from TNF-/- mice prevented GzmA-induced osteoclastogenesis. GzmA-deficient mice had reduced osteoclastogenesis in vivo (fewer calcitonin receptor-positive multinucleated cells and fewer transcripts for cathepsin K, matrix metalloproteinase 9, and TRAP in joints) and reduced serum levels of C-terminal telopeptide of type I collagen.Conclusion. GzmA contributes to the joint destruction of RA partly by promoting osteoclast differentiation.