Zhitao Rao1, Shuqing Wang2, Jiaqi Wang1. 1. Department of Orthopaedics, Tongji Hospital, Tongji University School of Medicine, Shanghai, 20065, China. 2. Department of Orthopaedics, Tongji Hospital, Tongji University School of Medicine, Shanghai, 20065, China. Electronic address: tongjijoint0106@163.com.
Abstract
BACKGROUND: Chondrocytes apoptosis induced by reactive oxygen species (ROS) plays a critical role in the pathogenesis of osteoarthritis (OA). Peroxiredoxin 4 (PRDX4), a member of the PRDX family, is essential for removing metabolic free radicals and reducing intracellular ROS. In this study, we sought to investigate the roles of PRDX4 on interleukin 1β (IL-1β)-induced chondrocyte apoptosis. METHODS: Primary chondrocytes were isolated from the articular cartilage of Sprague-Dawley rats, infected with PRDX4 overexpressing lentivirus and treated with IL-1β (10ng/mL). Cell apoptosis and ROS production identified by flow cytometry. Protein expression levels was evaluated by Western blotting analysis. Nitric oxide (NO) production and Caspase-3/9 activation were assessed by the Griess reaction method and colorimetric assay kit, respectively. RESULTS: PRDX4 overexpression in chondrocytes significantly decreased IL-1β-induced apoptosis. It also reversed the activity of IL-1β that increased ROS and NO production. PRDX4 overexpression reversed the activity of IL-1β that reduced the levels of Bcl-2, p-AKT and p-PRAS40, as well as increased Bax levels and Caspase-3/9 activation. More importantly, pre-treated with AKT inhibitor (AZD5363) significantly reduced the protective effects of PRDX4. CONCLUSIONS: Our data demonstrated that the regulatory effects of PRDX4 on IL-1β-induced chondrocyte apoptosis can be partially attributed to phosphatidylinositol 3-kinase/AKT signaling. These results indicate that PRDX4 might play a protective role in OA cartilage degeneration.
BACKGROUND: Chondrocytes apoptosis induced by reactive oxygen species (ROS) plays a critical role in the pathogenesis of osteoarthritis (OA). Peroxiredoxin 4 (PRDX4), a member of the PRDX family, is essential for removing metabolic free radicals and reducing intracellular ROS. In this study, we sought to investigate the roles of PRDX4 on interleukin 1β (IL-1β)-induced chondrocyte apoptosis. METHODS: Primary chondrocytes were isolated from the articular cartilage of Sprague-Dawley rats, infected with PRDX4 overexpressing lentivirus and treated with IL-1β (10ng/mL). Cell apoptosis and ROS production identified by flow cytometry. Protein expression levels was evaluated by Western blotting analysis. Nitric oxide (NO) production and Caspase-3/9 activation were assessed by the Griess reaction method and colorimetric assay kit, respectively. RESULTS:PRDX4 overexpression in chondrocytes significantly decreased IL-1β-induced apoptosis. It also reversed the activity of IL-1β that increased ROS and NO production. PRDX4 overexpression reversed the activity of IL-1β that reduced the levels of Bcl-2, p-AKT and p-PRAS40, as well as increased Bax levels and Caspase-3/9 activation. More importantly, pre-treated with AKT inhibitor (AZD5363) significantly reduced the protective effects of PRDX4. CONCLUSIONS: Our data demonstrated that the regulatory effects of PRDX4 on IL-1β-induced chondrocyte apoptosis can be partially attributed to phosphatidylinositol 3-kinase/AKT signaling. These results indicate that PRDX4 might play a protective role in OA cartilage degeneration.
Authors: Shi Sirong; Chen Yang; Tian Taoran; Li Songhang; Lin Shiyu; Zhang Yuxin; Shao Xiaoru; Zhang Tao; Lin Yunfeng; Cai Xiaoxiao Journal: Bone Res Date: 2020-02-10 Impact factor: 13.567
Authors: Simone Lipinski; Steffen Pfeuffer; Philipp Arnold; Christian Treitz; Konrad Aden; Henriette Ebsen; Maren Falk-Paulsen; Nicolas Gisch; Antonella Fazio; Jan Kuiper; Anne Luzius; Susanne Billmann-Born; Stefan Schreiber; Gabriel Nuñez; Hans-Dietmar Beer; Till Strowig; Mohamed Lamkanfi; Andreas Tholey; Philip Rosenstiel Journal: EMBO J Date: 2019-09-23 Impact factor: 11.598
Authors: Shi Sirong; Chen Yang; Tian Taoran; Li Songhang; Lin Shiyu; Zhang Yuxin; Shao Xiaoru; Zhang Tao; Lin Yunfeng; Cai Xiaoxiao Journal: Bone Res Date: 2020-02-10 Impact factor: 13.567