Abstract：
The imbalance between apoptosis and proliferation in fibroblast-like synoviocytes (FLSs) plays a key role in the pathogenesis of rheumatoid arthritis (RA). This study aims to investigate the potential of all-trans retinoic acid (ATRA) as a supplementary therapeutic agent alongside methotrexate (MTX) for RA, by examining its ability to inhibit synovial cell proliferation and enhance apoptosis through the ROS-JNK signalling pathway.
The viability, apoptosis, and autophagy levels of human rheumatoid arthritis fibroblast-like synovial cells (HFLS-RA) were evaluated, while ROS generation was measured through the DCFH-DA fluorescence microplate assay. Western blotting was used to analyse the expression levels of JNK signalling pathway-related proteins. To assess therapeutic potential in vivo, a collagen-induced arthritis (CIA) model was established in Wistar rats.
Small doses of MTX did not significantly affect the viability of HFLS-RAs or induce apoptosis. However, when ATRA was added to the treatment, the therapy markedly inhibited cell proliferation and induced apoptosis and excessive autophagy. Mechanistically, ATRA activated the ROS/JNK signalling pathway in HFLS-RAs. ROS scavengers and JNK inhibitors significantly attenuated ATRA-induced apoptosis and autophagy. In vivo, the combination therapy demonstrated a remarkable enhancement of the anti-arthritic efficacy in CIA rats.
The ability of ATRA to inhibit proliferation in RA FLSs through autophagy and apoptosis underscores its potential as a supplementary therapeutic agent alongside MTX for RA, particularly when compared to the limited impact of MTX on these processes. This combined strategy holds promise for enhancing therapeutic outcomes and warrants further investigation in the management of RA.
The viability, apoptosis, and autophagy levels of human rheumatoid arthritis fibroblast-like synovial cells (HFLS-RA) were evaluated, while ROS generation was measured through the DCFH-DA fluorescence microplate assay. Western blotting was used to analyse the expression levels of JNK signalling pathway-related proteins. To assess therapeutic potential in vivo, a collagen-induced arthritis (CIA) model was established in Wistar rats.
Small doses of MTX did not significantly affect the viability of HFLS-RAs or induce apoptosis. However, when ATRA was added to the treatment, the therapy markedly inhibited cell proliferation and induced apoptosis and excessive autophagy. Mechanistically, ATRA activated the ROS/JNK signalling pathway in HFLS-RAs. ROS scavengers and JNK inhibitors significantly attenuated ATRA-induced apoptosis and autophagy. In vivo, the combination therapy demonstrated a remarkable enhancement of the anti-arthritic efficacy in CIA rats.
The ability of ATRA to inhibit proliferation in RA FLSs through autophagy and apoptosis underscores its potential as a supplementary therapeutic agent alongside MTX for RA, particularly when compared to the limited impact of MTX on these processes. This combined strategy holds promise for enhancing therapeutic outcomes and warrants further investigation in the management of RA.
摘要：
目的：成纤维样滑膜细胞（FLSs）凋亡与增殖的失衡在类风湿关节炎（RA）的发病机制中起关键作用。本研究旨在探讨全反式维甲酸（ATRA）作为辅助治疗药物与甲氨蝶呤（MTX）治疗RA的潜力。通过ROS-JNK信号通路检测其抑制滑膜细胞增殖和增强细胞凋亡的能力。
方法：生存能力，凋亡，和自噬水平的人类风湿性关节炎成纤维细胞样滑膜细胞（HFLS-RA）进行了评估，而ROS的产生是通过DCFH-DA荧光微板测定来测量的。Western印迹用于分析JNK信号通路相关蛋白的表达水平。为了评估体内的治疗潜力，在Wistar大鼠中建立胶原诱导的关节炎（CIA）模型。
结果：小剂量的MTX没有显著影响HFLS-RAs的活力或诱导细胞凋亡。然而,当ATRA被添加到治疗中时，该疗法明显抑制细胞增殖，诱导细胞凋亡和过度自噬。机械上,ATRA激活了HFLS-RAs中的ROS/JNK信号通路。ROS清除剂和JNK抑制剂显著减弱ATRA诱导的细胞凋亡和自噬。在体内,联合治疗在CIA大鼠中显示出抗关节炎功效的显着增强。
结论：ATRA通过自噬和凋亡抑制RAFLS增殖的能力强调了其作为与MTX一起治疗RA的补充治疗剂的潜力，特别是与MTX对这些过程的有限影响相比。这种联合策略有望提高治疗效果，并需要对RA的管理进行进一步研究。
方法：生存能力，凋亡，和自噬水平的人类风湿性关节炎成纤维细胞样滑膜细胞（HFLS-RA）进行了评估，而ROS的产生是通过DCFH-DA荧光微板测定来测量的。Western印迹用于分析JNK信号通路相关蛋白的表达水平。为了评估体内的治疗潜力，在Wistar大鼠中建立胶原诱导的关节炎（CIA）模型。
结果：小剂量的MTX没有显著影响HFLS-RAs的活力或诱导细胞凋亡。然而,当ATRA被添加到治疗中时，该疗法明显抑制细胞增殖，诱导细胞凋亡和过度自噬。机械上,ATRA激活了HFLS-RAs中的ROS/JNK信号通路。ROS清除剂和JNK抑制剂显著减弱ATRA诱导的细胞凋亡和自噬。在体内,联合治疗在CIA大鼠中显示出抗关节炎功效的显着增强。
结论：ATRA通过自噬和凋亡抑制RAFLS增殖的能力强调了其作为与MTX一起治疗RA的补充治疗剂的潜力，特别是与MTX对这些过程的有限影响相比。这种联合策略有望提高治疗效果，并需要对RA的管理进行进一步研究。
