BRD-K20733377 may inhibit through the STAT3/NFKB1 axis, thereby alleviating (IVDD).
Thirty genes related to ferroptosis were identified as potentially linked to BRD-K20733377 and IVDD.
Core targets identified include STAT3 and NFKB1, which are validated as having a direct causal relationship with IVDD.
In vitro experiments indicated that BRD-K20733377 reduces intracellular iron levels and lipid peroxidation, regulating ferroptosis.
Ferroptosis-related proteins supported the proposed mechanism of BRD-K20733377 in inhibiting ferroptosis.
In vivo experiments demonstrated significant improvements in pain scores and behavior in rats treated with BRD-K20733377.
Simplified
INTRODUCTION: (IVDD) is one of the main causes of low back pain. Existing literature has confirmed the occurrence of 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.
Key numbers
< 0.05
Pain Score Reduction
Compared to the PBS group in the Von Frey test.
-7.0 kcal/mol
Binding Energy with
From molecular docking analysis of BRD with .
-7.4 kcal/mol
Binding Energy with
From molecular docking analysis of BRD with .
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Declarations. Ethics approval: The study titled “BRD-K20733377 inhibits ferroptosis and alleviates intervertebral disc degeneration via the STAT3/NFKB1 axis: a multiapproach study” was approved by the Animal Ethics Committee of Yangzhou University (Approval No. 202403237) on March 15, 2024. Additionally, the study protocol was approved by the Ethics Committee of the Northern Jiangsu People’s Hospital Affiliated to Yangzhou University (Approval No. 2021ky050) on March 2, 2021. Consent to participate: Informed consent was obtained from all individual participants included in the study. This study involving human NP tissue samples was approved by the Ethics Committee of the Northern Jiangsu People’s Hospital Affiliated to Yangzhou University (Approval No. 2021ky050) on March 2, 2021, under the project title: ‘BRD-K20733377 inhibits ferroptosis and alleviates intervertebral disc degeneration via the STAT3/NFKB1 axis: a multiapproach study’. Consent for publication: Our manuscript does not contain any individual person’s data in any form (including any individual details, images or videos). Competing interests: The authors declare no competing interests. AI use declaration: The authors declare that they have not use AI-generated work in this manuscript.
