白细胞介素-13 通过上调成纤维样滑膜细胞护骨素表达减少类风湿关节炎骨侵蚀：一项体外和体内研究。

Interleukin-13 reduces bone erosion in rheumatoid arthritis by up-regulating osteoprotegerin expression in fibroblast-like synoviocytes : an in vitro and in vivo study.

机构信息

Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, China.

Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, Clinical College of Nanjing Medical University, Nanjing, China.

出版信息

Clin Exp Rheumatol. 2023 Nov;41(11):2151-2161. doi: 10.55563/clinexprheumatol/b96n1e. Epub 2023 Mar 27.

DOI:10.55563/clinexprheumatol/b96n1e

PMID:36995338

Abstract

OBJECTIVES

Bone erosion in rheumatoid arthritis (RA) is partly caused by excessive activation of osteoclasts. Osteoclasts can be derived from RA synovium and their differentiation can be inhibited by osteoprotegerin (OPG), a decoy receptor of the osteoclastogenesis-promoting cytokine receptor activator of nuclear factor κB ligand (RANKL). Fibroblast-like synoviocytes (FLSs) are the main stromal cells in the synovium that can secret OPG. The OPG secretion of FLSs can be modulated by various cytokines. Interleukin (IL)-13 can alleviate bone erosion in RA mouse models, but the mechanisms remain unclear. Therefore, we aimed to investigate whether IL-13 can induce OPG secretion by RA-FLSs, thus ameliorating bone destruction in RA by inhibiting osteoclast differentiation.

METHODS

OPG, RANKL, and IL-13 receptors expression by RA-FLSs were evaluated by RT-qPCR. OPG secretion was determined by ELISA. Western blot was performed to analyse OPG expression and the activation of the STAT6 pathway. IL-13 and (or) OPG siRNA pre-treated RA-FLSs conditioned medium were used in osteoclast induction to test if IL-13 can inhibit osteoclastogenesis by up-regulating OPG in RA-FLSs. Micro-CT and immunofluorescence were performed to determine if IL-13 can induce OPG expression and alleviate bone erosion in vivo.

RESULTS

IL-13 can promote OPG expression of RA-FLSs, and the promotion can be overcome by IL-13Rα1 or IL-13Rα2 siRNA transfection, or STAT6 inhibitor. Osteoclast differentiation can be inhibited by IL-13 pre-treated RA-FLSs conditioned medium. The inhibition can be reversed by OPG siRNA transfection. IL-13 injection can increase OPG expression in the joints while reducing bone destruction in collagen-induced arthritis mice.

CONCLUSIONS

IL-13 can inhibit osteoclastogenesis by up-regulating OPG in RA-FLSs through IL-13 receptors via the STAT6 pathway, thus may ameliorate bone erosion in RA.

摘要

目的

类风湿关节炎（RA）中的骨侵蚀部分是由破骨细胞过度激活引起的。破骨细胞可以来源于 RA 滑膜，其分化可以被核因子κB 配体（RANKL）的破骨细胞生成促进细胞因子受体激活剂的诱饵受体骨保护素（OPG）抑制。成纤维样滑膜细胞（FLS）是滑膜中的主要基质细胞，可分泌 OPG。FLS 分泌的 OPG 可以被各种细胞因子调节。白细胞介素（IL）-13 可以减轻 RA 小鼠模型中的骨侵蚀，但机制尚不清楚。因此，我们旨在研究 IL-13 是否可以诱导 RA-FLS 分泌 OPG，从而通过抑制破骨细胞分化来改善 RA 中的骨破坏。

方法

通过 RT-qPCR 评估 RA-FLS 中 OPG、RANKL 和 IL-13 受体的表达。通过 ELISA 测定 OPG 分泌。通过 Western blot 分析 OPG 表达和 STAT6 通路的激活。使用 IL-13 和（或）OPG siRNA 预处理的 RA-FLS 条件培养基进行破骨细胞诱导，以测试 IL-13 是否可以通过上调 RA-FLS 中的 OPG 来抑制破骨细胞生成。通过 micro-CT 和免疫荧光测定，确定 IL-13 是否可以诱导 OPG 表达并减轻体内骨侵蚀。

结果

IL-13 可以促进 RA-FLS 中 OPG 的表达，这种促进作用可以通过 IL-13Rα1 或 IL-13Rα2 siRNA 转染或 STAT6 抑制剂来克服。IL-13 预处理的 RA-FLS 条件培养基可以抑制破骨细胞分化。这种抑制作用可以通过 OPG siRNA 转染逆转。IL-13 注射可以增加关节炎关节中的 OPG 表达，同时减少胶原诱导关节炎小鼠的骨破坏。