背景:骨关节炎(OA)是老年人群最常见的关节疾病之一。促炎细胞因子,如白细胞介素-1β(IL-1β),在OA的发展过程中发挥着重要作用。Dapansutrile是NOD样受体蛋白3(NLRP3)炎性体的特异性抑制剂,具有抗炎特性。
方法:在本研究中,我们研究了dapanstryle对软骨退变的保护作用及其机制。在本研究中,从大鼠中分离出软骨细胞,然后用达潘舒利治疗。之后,(Cox-2,诱导型一氧化氮合酶(iNOS),Mmp-3,Mmp-9,Mmp-13和IL-10)在RNA水平上进行评估,然后在蛋白质水平评估(COX-2,MMP-3,MMP-9,MMP-13,SOX-9和COL2)的表达。随后,使用蛋白质印迹法(WB)检测丝裂原活化蛋白激酶(MAPK)途径的活化.此外,建立了大鼠OA模型,以评估达潘舒利的体内保护作用。
结果:结果表明,dapansutrile在24h(0、1、2、5和10μM)对大鼠软骨细胞没有明显的细胞毒性。Dapansutrile显着降低IL-1β诱导的COX2,iNOS的上调,基质金属蛋白酶3(MMP3),9(MMP9)和13(MMP13),并逆转IL-1β诱导的IL-10、SOX9和COL2的下调。Dapansutrile还通过下调磷酸化ERK的表达水平来抑制IL-1β诱导的MAPK信号通路的上调,和磷酸-P38浓度依赖的方式。此外,dapansutrile在具有较低Mankin评分和国际骨关节炎研究协会(OARSI)评分的大鼠OA模型中表现出保护作用。
结论:我们的研究表明达巴苏曲在体外通过抑制MAPK信号通路有效抑制软骨细胞炎症。和改善体内软骨退化,表明在OA治疗中具有抗炎作用。
BACKGROUND: Osteoarthritis (OA) is one of the most common joint diseases in the elderly population. Proinflammatory cytokines, such as Interleukin-1β (IL-1β), play an important role in the development and progression of OA. Dapansutrile is a specific inhibitor of the NOD-like receptor protein 3 (NLRP3) inflammasome and exhibits anti-inflammatory properties.
METHODS: In this study, we investigated the protective effect and the underlying mechanism of dapansutrile on cartilage degeneration in vitro and in vivo. In the present study, chondrocytes were isolated from rats and then were treated with dapansutrile. After that, the expression of (Cox-2, inducible nitric oxide synthase (iNOS), Mmp-3, Mmp-9, Mmp-13 and IL-10) were evaluated at RNA level, then the expression of (COX-2, MMP-3, MMP-9, MMP-13, SOX-9 and COL2) were evaluated at protein level. Subsequently, the activation of the mitogen-activated protein kinase (MAPK) pathway was tested using western blotting (WB). Additionally, the rat OA model was developed to evaluate the protective effects of dapansutrile in vivo.
RESULTS: The results showed that dapansutrile had no obvious cytotoxicity on rat chondrocytes at 24 h (0, 1, 2, 5 and 10 μM). Dapansutrile significantly decreased IL-1β-induced upregulation of COX2, iNOS, matrix metalloproteinase 3 (MMP3), 9 (MMP9) and 13 (MMP13), and reversed IL-1β-induced the downregulation of IL-10, SOX9 and COL2. Dapansutrile also inhibited IL-1β-induced upregulation of the MAPK signaling pathway by downregulating the expression levels of phospho-ERK, and phospho-P38 in a concentration dependent manner. In addition, dapansutrile exhibited protective effects in rat OA model with lower Mankin\'s score and Osteoarthritis Research Society International (OARSI) score.
CONCLUSIONS: Our study suggested that dapansutrile effectively inhibited chondrocyte inflammation by suppressing MAPK signaling pathway in vitro, and ameliorated cartilage degeneration in vivo, indicating an anti-inflammatory effect in OA treatment.