Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, UK.
Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, UK.
Clin Exp Rheumatol. 2022 Sep;40(9):1620-1628. doi: 10.55563/clinexprheumatol/cfsajk. Epub 2021 Oct 13.
To investigate the ex vivo effect of the JAK1/2 inhibitor baricitinib on expression of pro-inflammatory mediators in rheumatoid arthritis (RA) fibroblast like synoviocytes (FLS) stimulated with TNFα, IL-1β and oncostatin M (OSM), and in RA synovial membrane cells (SMCs).
RA and osteoarthritis (OA) SMCs, were isolated from arthroplasty specimens of RA (n=8) and OA (n=8) patients, respectively, using enzymatic digestion followed by cell propagation to obtain RA (n=5) and OA (n=3) FLS. Normal FLS and normal human foreskin fibroblasts (HSF) were purchased from commercial sources. Fibroblasts were stimulated with cytokines with or without baricitinib. RA SMCs were cultured in the presence of baricitinib without stimulation. JAK/STAT activation and levels of mRNA and proteins of the various inflammatory cytokines (IL-6, IL-8, MCP-1, RANTES and IP-10) were determined by qPCR, ELISA and MSD.
Baricitinib inhibited OSM-induced JAK signalling in RA synovial fibroblasts and effectively suppressed subsequent expression of the proinflammatory mediators IL-6, MCP-1 and IP-10. However, baricitinib was not effective in altering levels of spontaneously released TNFα, IL-6 and IL-8 in RA SMC. Although both TNFα and IL-1β signal independently of the JAK/STAT pathway, in HSF, but not in RA FLS, baricitinib significantly inhibited TNFα- and IL-1β-induced MCP-1 and IP-10 protein levels in a dose dependent manner. Furthermore, baricitinib did not inhibit TNFα- and IL-1β-induced expression of IL-6, IL-8 and MCP-1 in RA FLS.
These findings are consistent with known signalling pathways employed by OSM, TNFα and IL-1β, but our data suggest that in HSF, baricitinib may have anti-inflammatory effects via downstream modulation of cytokines and chemokines produced in response to TNFα or IL-1β.
研究 JAK1/2 抑制剂巴瑞替尼对肿瘤坏死因子 α（TNFα）、白细胞介素 1β（IL-1β）和抑瘤素 M（OSM）刺激的类风湿关节炎（RA）成纤维样滑膜细胞（FLS）中促炎介质表达的体外作用，以及在 RA 滑膜细胞（SMCs）中的作用。
RA 和骨关节炎（OA）SMC 分别从 RA（n=8）和 OA（n=8）患者的关节置换标本中通过酶消化和细胞传代获得，用 RA（n=5）和 OA（n=3）FLS 进行培养。正常 FLS 和正常人包皮成纤维细胞（HSF）购自商业来源。用细胞因子刺激成纤维细胞，或用巴瑞替尼与细胞因子共同刺激。RA SMC 在无刺激的情况下用巴瑞替尼培养。通过 qPCR、ELISA 和 MSD 测定 JAK/STAT 激活和各种炎症细胞因子（IL-6、IL-8、MCP-1、RANTES 和 IP-10）的 mRNA 和蛋白水平。
巴瑞替尼抑制 RA 滑膜成纤维细胞中 OSM 诱导的 JAK 信号转导，并有效抑制随后促炎介质 IL-6、MCP-1 和 IP-10 的表达。然而，巴瑞替尼不能改变 RA SMC 中自发释放的 TNFα、IL-6 和 IL-8 的水平。虽然 TNFα和 IL-1β独立于 JAK/STAT 途径信号，但在 HSF 中，而不是在 RA FLS 中，巴瑞替尼以剂量依赖性方式显著抑制 TNFα和 IL-1β诱导的 MCP-1 和 IP-10 蛋白水平。此外，巴瑞替尼不能抑制 RA FLS 中 TNFα和 IL-1β诱导的 IL-6、IL-8 和 MCP-1 的表达。
这些发现与 OSM、TNFα 和 IL-1β 所采用的已知信号通路一致，但我们的数据表明，在 HSF 中，巴瑞替尼可能通过下游调节 TNFα 或 IL-1β 反应性细胞因子和趋化因子的表达发挥抗炎作用。
