INTRODUCTION:Intervertebral disc degeneration (IVDD) is one of the main causes of low back pain. Existing literature has confirmed the occurrence of ferroptosis in IVDD. With the widespread application of artificial intelligence (AI), an increasing number of compounds have been screened for their potential to alleviate IVDD. BRD-K20733377 is one such compound with anti-aging properties. Preliminary experiments have shown that BRD-K20733377 can significantly inhibit cellular ferroptosis. However, research on the potential therapeutic targets and mechanisms of BRD-K20733377 in IVDD remains limited.
OBJECTIVE:This study aims to explore the main targets and potential mechanisms of BRD-K20733377 in the treatment of IVDD.
METHODS:Network pharmacology, bioinformatics, machine learning algorithms, molecular docking, molecular dynamics, and Mendelian randomization were used to comprehensively analyze the effects, potential targets, and mechanisms of BRD-K20733377 in IVDD. Rat nucleus pulposus-derived mesenchymal stem cells (NPMSCs) were selected for in vitro experiments. Cell viability was measured by CCK-8 and flow cytometry, while antioxidant defense, lipid peroxidation, and iron metabolism were explored using JC-1, Reactive Oxygen Species (ROS), FerroOrange dye, Lipid peroxides, Malondialdehyde (MDA), and Mitochondrial transmission electron microscopy. The expression levels of ferroptosis-related proteins were detected by Western blot and immunofluorescence. In the rat IVDD model, the effects of BRD-K20733377 on pain threshold, pain behavior, and its therapeutic efficacy were evaluated.
RESULTS:Predictive results identified 30 genes related to ferroptosis in BRD-K20733377 and IVDD, revealing that the potential mechanism of BRD-K20733377 in treating IVDD is closely associated with ferroptosis. In addition, functional enrichment analysis indicated that these genes are involved in multiple signaling pathways. Machine learning algorithms further identified the core targets STAT3 and NFKB1, and Mendelian randomization validated their direct causal relationship with IVDD. In vitro experiments confirmed that BRD-K20733377 inhibited IVDD by reducing intracellular Fe²⁺ levels and lipid peroxidation, thus regulating ferroptosis. Theoretically, BRD-K20733377 may inhibit NPMSCs ferroptosis via STAT3/NFKB1 axis. Ferroptosis-related proteins and immunofluorescence results further supported this mechanism. In vivo experiments showed that BRD-K20733377 significantly improved the behavior of SD rats, reduced pain scores, and alleviated IVDD.
CONCLUSION:BRD-K20733377 inhibits ferroptosis through the STAT3/NFKB1 axis, thereby alleviating IVDD. This provides a new perspective for the study of IVDD and could serve as a potential therapeutictarget for IVDD.
