Duhuo Jisheng Decoction suppresses apoptosis and mitochondrial dysfunction in human nucleus pulposus cells by miR-494/SIRT3/mitophagy signal axis

• Published in: Journal of orthopaedic surgery and research 2023-03, Vol.18 (1), p.177-177, Article 177
• Main Authors: Liu, Wei, Zhao, Xiaolong, Wu, Xuejian
• Format: Article
• Language: English
• Subjects: Apoptosis; Arthritis; Autophagy; Cells, Cultured; Cyclosporin A; Degeneration; Duhuo Jisheng Decoction; Extracellular matrix; Homeostasis; Humans; Hybridization; IL-1β; Intervertebral Disc Degeneration - pathology; Intervertebral discs; Intervertebral disk degeneration; Medicine, Chinese; MicroRNAs - metabolism; miR-494; Mitochondria; Mitochondria - genetics; Mitophagy; Nucleus Pulposus; Orthopedics; Osteoarthritis; Senescence; Signal transduction; SIRT3; Sirtuin 3 - genetics; Sirtuin 3 - metabolism; Traditional Chinese medicine; Western blotting
• ISSN: 1749-799X; 1749-799X
• DOI: 10.1186/s13018-023-03669-w

Increasing evidence suggests that mitophagy is responsible for the pathogenesis of intervertebral disk (IVD) degeneration. Previous studies have shown that Duhuo Jisheng Decoction (DHJSD), a classic Fangji of traditional Chinese medicine, can delay IVD degeneration; however, its specific mechanism of action is unknown. In this study, we investigated the mechanism by which DHJSD treatment prevented IVD degeneration in IL-1β-treated human nucleus pulposus (NP) cells in vitro. Cell Counting Kit-8 was performed to explore the effects of DHJSD on the viability of NP cells exposed to IL-1β. The mechanism by which DHJSD delays IVD degeneration was explored using luciferase reporter assay, RT-qPCR, western blotting, TUNEL assay, mitophagy detection assay, Mito-SOX, Mitotracker and in situ hybridization. We observed that DHJSD enhanced the viability of NP cells treated with IL-1β in a concentration-time dependent approach. Moreover, DHJSD lessened IL-1β-induced NP apoptosis and mitochondrial dysfunction and activated mitophagy in NP cells treated with IL-1β. Mitophagy suppressor cyclosporin A reversed the beneficial impacts of DHJSD in NP cells. In addition, the differential expression of miR-494 regulated IL-1β-induced NP apoptosis and mitochondrial dysfunction, and the protective impact of miR-494 on NP cells treated with IL-1β was achieved by mitophagy activation, which was regulated by its target gene, sirtuin 3 (SIRT3). Finally, we observed that DHJSD treatment could effectively delay IL-1β-induced NP apoptosis by affecting the miR-494/SIRT3/mitophagy signal axis. These results show that the miR-494/SIRT3/mitophagy signaling pathway is responsible for the apoptosis and mitochondrial dysfunction of NP cells and that DHJSD may exert protective effects against IVD degeneration by regulating the miR-494/SIRT3/mitophagy signal axis.