Nrf-2/ROS/NF-B pathway is modulated by cynarin in human mesenchymal stem cells in vitro from ankylosing spondylitis

Ankylosing spondylitis (AS) is an immune chronic inflammatory disease, resulting in back pain, stiffness, and thoracolumbar kyphotic deformity. Based on the reported anti-inflammatory and antioxidant capacities of cynarin (Cyn), this study explored its protective role and molecular mechanisms in mesenchymal stem cells (MSCs) from AS. The target pathways and genes were verified using Western blotting, quantitative real-time polymerase chain reaction, and immunofluorescent staining, while molecular docking analysis was conducted. In AS-MSCs, we found that the expression levels of p-NF-kappa B, IL-6, IL-1 beta, and TNF-alpha were higher and I kappa B-alpha, Nrf-2, and HO-1 were lower compared with healthy control (HC)-MSCs. With molecular docking analysis, the biding affinities between Cyn and Keap1-Nrf-2 and p65-I kappa B-alpha were predicted. The mRNA and protein expression of p-NF-kappa B, IL-6, IL-1 beta, and TNF-alpha and the reactive oxygen species (ROS) generation were downregulated following Cyn administration. Meanwhile, the expression level of I kappa B-alpha, Nrf-2, and HO-1 were significantly increased after Cyn pretreatment. The results suggested that the protective mechanisms of Cyn in AS-MSCs were based on enhancing the antioxidation and suppression of excessive inflammatory responses via Nrf-2/ROS/NF-kappa B axis. Our findings demonstrate that Cyn is a potential candidate for alleviating inflammation in AS.