Oliveira P G, Grespan R, Pinto L G, Meurer L, Brenol J C T, Roesler R, Schwartsmann G, Cunha F Q, Xavier R M
Hospital de Clinicas de Porto Alegre, Serviço de Reumatologia, and Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
Arthritis Rheum. 2011 Oct;63(10):2956-65. doi: 10.1002/art.30486.
To evaluate the antiinflammatory effects of RC-3095 in 2 experimental models of arthritis, collagen-induced arthritis (CIA) and antigen-induced arthritis (AIA), and to determine the mechanisms of action involved.
RC-3095 was administered daily to mice with CIA and mice with AIA, after induction of disease with methylated bovine serum albumin. Disease incidence and severity were assessed using a clinical index and evaluation of histologic features, respectively. In mice with CIA, gastrin-releasing peptide receptor (GRPR) was detected by immunohistochemical analysis, while in mice with AIA, migration of neutrophils, presence of glycosaminoglycans, and lymphocyte proliferation, determined using the MTT assay, were assessed. Expression of cytokines interleukin-17 (IL-17), IL-1β, and tumor necrosis factor α (TNFα) was evaluated in all mouse knees using enzyme-linked immunosorbent assay. Treg cell production was assessed by flow cytometry in the joints of mice with AIA.
In mice with AIA, administration of RC-3095 reduced neutrophil migration, mechanical hypernociception, and proteoglycan loss. These findings were associated with inhibition of the levels of all 3 proinflammatory cytokines, decreased lymphocyte proliferation, and increased Treg cell numbers. In the CIA model, treatment with RC-3095 led to a significant reduction in arthritis clinical scores and the severity of disease determined histologically. Synovial inflammation, synovial hyperplasia, pannus formation, and extensive erosive changes were all dramatically reduced in the arthritic mice treated with RC-3095. Furthermore, arthritic mice treated with RC-3095 showed a significant reduction in the concentrations of IL-17, IL-1β, and TNFα, and showed a diminished expression of GRPR.
These findings suggest that the GRP pathway has a significant role in chronic arthritis, and its inhibition can be explored as a possible therapeutic strategy in rheumatoid arthritis.
评估RC - 3095在两种实验性关节炎模型——胶原诱导性关节炎（CIA）和抗原诱导性关节炎（AIA）中的抗炎作用，并确定其作用机制。
在用甲基化牛血清白蛋白诱导疾病后，每天给患有CIA和AIA的小鼠施用RC - 3095。分别使用临床指标和组织学特征评估疾病发病率和严重程度。在患有CIA的小鼠中，通过免疫组织化学分析检测胃泌素释放肽受体（GRPR），而在患有AIA的小鼠中，使用MTT法评估中性粒细胞迁移、糖胺聚糖的存在以及淋巴细胞增殖情况。使用酶联免疫吸附测定法评估所有小鼠膝关节中细胞因子白细胞介素 - 17（IL - 17）、IL - 1β和肿瘤坏死因子α（TNFα）的表达。通过流式细胞术评估患有AIA的小鼠关节中的调节性T细胞（Treg）产生情况。
在患有AIA的小鼠中，施用RC - 3095可减少中性粒细胞迁移、机械性痛觉过敏和蛋白聚糖损失。这些发现与所有三种促炎细胞因子水平的抑制、淋巴细胞增殖减少以及Treg细胞数量增加有关。在CIA模型中，用RC - 3095治疗导致关节炎临床评分和组织学确定的疾病严重程度显著降低。在用RC - 3095治疗的关节炎小鼠中，滑膜炎症、滑膜增生、血管翳形成和广泛的侵蚀性变化均显著减少。此外，用RC - 3095治疗的关节炎小鼠中IL - 17、IL - 1β和TNFα浓度显著降低，并且GRPR表达减少。
这些发现表明GRP通路在慢性关节炎中起重要作用，其抑制可作为类风湿性关节炎的一种可能治疗策略进行探索。
