Abstract

Considered as an autoimmune disease, rheumatoid arthritis (RA) is an chronic inflammatory disorder that causes inflammation of the joints. This study is performed with the aim to clarify the expression of phospholipase D1 (PLD1) in RA and its specific regulation role of RA as well as the underlying mechanisms. In this study, synovial tissue samples were collected from RA patients, and RA-fibroblast-like synoviocytes (FLSs) were subsequently isolated. The expression levels of PLD1 and pathway-related proteins were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting or immunohistochemistry (IHC). Upon shPLD1 treatment, cell viability, proliferation, migration, invasion, and the level of inflammation-related factors were measured by Cell Counting Kit-8 (CCK-8), Edu, wound healing, Transwell and enzyme-linked immunosorbent assay (ELISA). Furthermore, C-reactive protein (CRP), rheumatoid factor (RF), arthritis score and synovial tissue lesions were assessed by collecting the blood or tissues from collagen induced arthritis (CIA) model rats. Our results showed that PLD1 level was increased in RA synovial tissues. Cell viability, proliferation, migration, invasion, and the level of inflammatory factors were reduced upon PLD1 knockdown in RA-FLSs. Moreover, p-IκBα/IκBα, β-catenin, p-IKKβ/IKKβ and TCF-4 were inhibited under PLD1 knockdown treatment. PLD1 knockdown alleviated the collagen-induced addition of arthritis score, CRP and RF, as well as the filling of inflammatory cells and proliferation of synovium in CIA model rat. To sum up, knockdown of PLD1 could reduce RA-FLSs metastasis as well as inflammatory response by modulating the activity of NF-κB and Wnt/β-catenin pathways.

摘要

类风湿性关节炎 (RA) 被认为是一种自身免疫性疾病，是一种导致关节炎症的慢性炎症性疾病。本研究旨在阐明磷脂酶 D1 (PLD1) 在 RA 中的表达及其对 RA 的特定调节作用及其潜在机制。在这项研究中，s从 RA 患者身上收集滑膜组织样本，随后分离出 RA 成纤维细胞样滑膜细胞 (FLS)。通过定量逆转录聚合酶链反应（qRT-PCR）、蛋白质印迹或免疫组织化学（IHC）检测PLD1和通路相关蛋白的表达水平。在 shPLD1 处理后，通过 Cell Counting Kit-8 (CCK-8)、Edu、伤口愈合、Transwell 和酶联免疫吸附试验 (ELISA) 测量细胞活力、增殖、迁移、侵袭和炎症相关因子的水平. 此外，通过收集胶原诱导的关节炎（CIA）模型大鼠的血液或组织，评估 C 反应蛋白（CRP）、类风湿因子（RF）、关节炎评分和滑膜组织损伤。我们的结果表明，RA 滑膜组织中 PLD1 水平升高。在 RA-FLS 中敲低 PLD1 后，细胞活力、增殖、迁移、侵袭和炎症因子水平降低。此外，p-IκBα/IκBα、β-连环蛋白、p-IKKβ/IKKβ 和 TCF-4 在 PLD1 敲低处理下受到抑制。PLD1 敲低减轻了胶原诱导的关节炎评分、CRP 和 RF 的增加，以及 CIA 模型大鼠中炎症细胞的填充和滑膜的增殖。综上所述，PLD1 的敲低可以通过调节 NF-κB 和 Wnt/β-catenin 通路的活性来减少 RA-FLSs 的转移以及炎症反应。PLD1 敲低减轻了胶原诱导的关节炎评分、CRP 和 RF 的增加，以及 CIA 模型大鼠中炎症细胞的填充和滑膜的增殖。综上所述，PLD1 的敲低可以通过调节 NF-κB 和 Wnt/β-catenin 通路的活性来减少 RA-FLSs 的转移以及炎症反应。PLD1 敲低减轻了胶原诱导的关节炎评分、CRP 和 RF 的增加，以及 CIA 模型大鼠中炎症细胞的填充和滑膜的增殖。综上所述，PLD1 的敲低可以通过调节 NF-κB 和 Wnt/β-catenin 通路的活性来减少 RA-FLSs 的转移以及炎症反应。