Kucharska-Lusina Aleksandra, Skrzypek Maciej, Tokarczyk Agnieszka, Dragan Grzegorz, Majsterek Ireneusz
Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, 92-215 Lodz, Poland.
Int J Mol Sci. 2025 Mar 11;26(6):2489. doi: 10.3390/ijms26062489.
Rheumatoid arthritis (RA) is a chronic, common autoimmune disease. It is characterized by inflammatory polyarthritis, which can lead to permanent disability in patients. Current treatment is mainly symptom-related, aiming to reduce pain and inflammation, but does not lead to a full recovery. This treatment includes non-steroidal anti-inflammatory drugs (NSAIDs) and disease-modifying anti-rheumatic drugs (DMARDs). It has been shown that, due to chronic inflammation, reduced glucose levels and hypoxia, endoplasmic reticulum (ER) stress is induced in RA patients, leading to the activation of multiple signaling pathways, including the ER-dependent adaptation of the unfolded protein response (UPR) pathway. The aim of this study was to assess the level of apoptosis in patients diagnosed with RA. The study sought to investigate whether UPR response correlated with apoptosis induction could serve as a potential diagnostic marker or therapeutic target. In vitro studies have shown that UPR pathway activity can be observed in patients diagnosed with RA. The study group consisted of PBMC cells from 61 individuals, including a total of 31 rheumatoid arthritis patients and 30 healthy controls. In order to validate UPR activation, we estimated molecular markers of ER stress via RT-qPCR expression analysis. expression was used as a standard control. Elevated levels of mRNA for the (-value = 0.001903), the () (-value = 0.007457 × 10) and the (-value = 0.002212) were confirmed in a group of RA patients. Further analysis showed that after the induction of apoptosis the percentage of DNA contained in the tail was 37.78% higher in RA patients than in the control group (-value = 0.0003) measured by comet assay. The exogenous damage caused by hydrogen peroxide was found to be statistically elevated in RA patients and the caspase-3 level was calculated of 40.17% higher than in controls (-value = 0.0028). It was also found that PBMC cells from RA patients were more sensitive to apoptotic induction. Our results were confirmed by flow cytometry. The most important finding from our data was the confirmation of elevated sensitivity to apoptosis induction in RA patients; the results showed a 40.23% higher percentage of cells in early apoptosis than in the control group (-value = 0.0105). Our results may help to assess the feasibility of the application of early diagnosis and targeted therapy in the treatment of RA patients, including the ER signaling pathway via selected UPR-dependent molecular inhibitors.
类风湿关节炎（RA）是一种慢性常见的自身免疫性疾病。其特征为炎症性多关节炎，可导致患者永久性残疾。目前的治疗主要针对症状，旨在减轻疼痛和炎症，但无法实现完全康复。这种治疗包括非甾体抗炎药（NSAIDs）和改善病情抗风湿药（DMARDs）。研究表明，由于慢性炎症、血糖水平降低和缺氧，类风湿关节炎患者会出现内质网（ER）应激，从而导致多种信号通路激活，包括未折叠蛋白反应（UPR）途径的内质网依赖性适应性反应。本研究旨在评估类风湿关节炎患者的细胞凋亡水平。该研究试图探究与细胞凋亡诱导相关的UPR反应是否可作为潜在的诊断标志物或治疗靶点。体外研究表明，在类风湿关节炎患者中可观察到UPR途径的活性。研究组由61名个体的外周血单核细胞（PBMC）组成，其中包括31名类风湿关节炎患者和30名健康对照。为验证UPR激活，我们通过逆转录定量聚合酶链反应（RT-qPCR）表达分析估计内质网应激的分子标志物。 表达用作标准对照。在一组类风湿关节炎患者中，确认 （-值 = 0.001903）、 （）（-值 = 0.007457 × 10）和 （-值 = 0.002212）的mRNA水平升高。进一步分析表明，通过彗星试验测量，在诱导细胞凋亡后，类风湿关节炎患者尾部所含DNA的百分比比对照组高37.78%（-值 = 0.0003）。发现过氧化氢引起的外源性损伤在类风湿关节炎患者中具有统计学意义的升高，且半胱天冬酶 - 3水平计算得出比对照组高40.17%（-值 = 0.0028）。还发现类风湿关节炎患者的PBMC细胞对细胞凋亡诱导更敏感。我们的结果通过流式细胞术得到证实。我们数据中最重要的发现是确认类风湿关节炎患者对细胞凋亡诱导的敏感性升高；结果显示早期凋亡细胞的百分比比对照组高40.23%（-值 = 0.0105）。我们的结果可能有助于评估在类风湿关节炎患者治疗中应用早期诊断和靶向治疗的可行性，包括通过选定的UPR依赖性分子抑制剂作用于内质网信号通路。
