Beijing University of Chinese Medicine, China-Japan Friendship Hospital Clinical Medicine, Beijing, China.
Institute of Clinical Medicine, China-Japan Friendship Hospital, Beijing, China.
Front Immunol. 2023 Mar 13;14:1135014. doi: 10.3389/fimmu.2023.1135014. eCollection 2023.
Necroptosis has recently been found to be associated with the pathogenesis of many autoimmune diseases, including rheumatoid arthritis (RA). This study was undertaken to explore the role of RIPK1-dependent necroptosis in the pathogenesis of RA and the potential new treatment options.
The plasma levels of receptor-interacting protein kinase 1 (RIPK1) and mixed lineage kinase domain-like pseudokinase (MLKL) in 23 controls and 42 RA patients were detected by ELISA. Collagen-induced arthritis (CIA) rats were treated with KW2449 by gavage for 28 days. Arthritis index score, H&E staining, and Micro-CT analysis were used to evaluate joint inflammation. The levels of RIPK1-dependent necroptosis related proteins and inflammatory cytokines were detected by qRT-PCR, ELISA and Western blot, and the cell death morphology was detected by flow cytometry analysis and high-content imaging analysis.
The plasma levels of RIPK1 and MLKL in RA patients were higher than those in healthy people, and were positively correlated with the severity of RA. KW2449 could reduce joint swelling, joint bone destruction, tissue damage, and the plasma levels of inflammatory cytokines in CIA rats. Lipopolysaccharide combined with zVAD (LZ) could induce necroptosis in RAW 264.7 cells, which could be reduced by KW2449. RIPK1-dependent necroptosis related proteins and inflammatory factors increased after LZ induction and decreased after KW2449 treatment or knockdown of RIPK1.
These findings suggest that the overexpression of RIPK1 is positively correlated with the severity of RA. KW2449, as a small molecule inhibitor targeting RIPK1, has the potential to be a therapeutic strategy for RA treatment by inhibiting RIPK1-dependent necroptosis.
坏死性凋亡最近被发现与许多自身免疫性疾病的发病机制有关，包括类风湿关节炎（RA）。本研究旨在探讨 RIPK1 依赖性坏死性凋亡在 RA 发病机制中的作用及潜在的新治疗选择。
采用 ELISA 法检测 23 例对照和 42 例 RA 患者的受体相互作用蛋白激酶 1（RIPK1）和混合谱系激酶结构域样伪激酶（MLKL）的血浆水平。采用灌胃 KW2449 的方法对胶原诱导性关节炎（CIA）大鼠进行 28 天治疗。采用关节炎指数评分、H&E 染色和 Micro-CT 分析评估关节炎症。采用 qRT-PCR、ELISA 和 Western blot 检测 RIPK1 依赖性坏死性凋亡相关蛋白和炎症细胞因子水平，采用流式细胞术分析和高内涵成像分析检测细胞死亡形态。
RA 患者的血浆 RIPK1 和 MLKL 水平高于健康人，且与 RA 的严重程度呈正相关。KW2449 可减轻 CIA 大鼠的关节肿胀、关节骨破坏、组织损伤和炎症细胞因子的血浆水平。脂多糖联合 zVAD（LZ）可诱导 RAW 264.7 细胞发生坏死性凋亡，KW2449 可减少这种凋亡。LZ 诱导后 RIPK1 依赖性坏死性凋亡相关蛋白和炎症因子增加，KW2449 处理或敲低 RIPK1 后减少。
这些发现表明 RIPK1 的过度表达与 RA 的严重程度呈正相关。KW2449 作为一种针对 RIPK1 的小分子抑制剂，通过抑制 RIPK1 依赖性坏死性凋亡，有可能成为治疗 RA 的一种治疗策略。
