成纤维细胞样滑膜细胞(FLSs),类风湿性关节炎(RA)的主要病理细胞,显示肿瘤样表型,包括过度扩散,凋亡抗性,和侵略性表型。RA-FLSs的过度增殖和凋亡不足可导致滑膜组织增生,炎症介质的过量产生,和关节的破坏。在这篇文章中,我们研究了PRIMA-1MET对RA-FLSs细胞凋亡诱导和促炎细胞因子抑制的影响。滑膜组织样品取自10例RA患者。用不同浓度的PRIMA-1MET处理FLSs。通过流式细胞术和MTT法评估细胞凋亡率和细胞存活率,并进行实时定量PCR以评估p53,IL-6,IL-1β的转录,TNF-α,Noxa,P21,PUMA,Bax,幸存者,和XIAP在治疗的RA-FLS中。p53、IκBα、和磷酸-IκBα使用蛋白质印迹法测量。结果表明,PRIMA-1MET诱导RA-FLSs细胞凋亡,并显著增加Noxa的表达,显著降低IL-6、IL-1β、p53和磷酸-IκBα表达。PRIMA-1MET可以通过诱导Noxa表达而下调p53来诱导RA-FLSs的凋亡。此外,PRIMA-1MET治疗导致促炎细胞因子产生的抑制和NF-κB抑制。鉴于p53和NF-κB在RA-FLS中的作用,PRIMA-1MET可以被认为是类风湿关节炎的一种新的治疗策略。
Fibroblast-like synoviocytes (FLSs), the main pathological cells in rheumatoid arthritis (RA), display tumor-like phenotype, including hyper-proliferation, apoptosis resistance, and aggressive phenotype. Excessive proliferation and insufficient apoptosis of RA-FLSs can lead to hyperplastic synovial pannus tissue, excess production of inflammatory mediators, and destruction of joints. In this article, we investigate the effect of PRIMA-1MET on the apoptosis induction and inhibition of pro-inflammatory cytokines in RA-FLSs. Synovial tissue samples were obtained from 10 patients with RA. The FLSs were treated with different concentrations of PRIMA-1MET. The rate of apoptosis and cell survival was assessed by flow cytometry and MTT assay and Real-time quantitative PCR was performed to evaluate the transcription of p53, IL-6, IL-1β, TNF-α,
Noxa, p21, PUMA, Bax, Survivin, and XIAP in treated RA-FLSs. The protein level of p53, IκBα, and phospho-IκBα were measured using Western blotting. The results showed that PRIMA-1MET induced apoptosis in RA-FLSs and increased significantly the expression of
Noxa, and decreased significantly IL-6, IL-1β, p53, and phospho-IκBα expression. PRIMA-1MET can induce apoptosis in RA-FLSs through induction of
Noxa expression while p53 was downregulated. Furthermore, PRIMA-1MET treatment results in the suppression of pro-inflammatory cytokine production and NF-κB inhibition. Given the role of p53 and NF-κB in RA-FLSs, PRIMA-1MET can be considered as a new therapeutic strategy for rheumatoid arthritis.