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BRD9 Inhibition Attenuates Matrix Degradation and Pyroptosis in Nucleus Pulposus by Modulating the NOX1/ROS/NF-κB axis

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Abstract

Intervertebral disc degeneration (IDD) is considered to be the leading cause of low back pain (LBP). The progression of IDD is closely related to the inflammatory microenvironment, which results in extracellular matrix degradation and cell death. One of the proteins, which have been shown to participate in the inflammatory response, is the bromodomain-containing protein 9 (BRD9). This study aimed to investigate the role and mechanism of BRD9 in regulating IDD. The tumor necrosis factor-α (TNF-α) was used to mimic the inflammatory microenvironment in vitro. Western blot, RT-PCR, immunohistochemistry, immunofluorescence, and flow cytometry were used to demonstrate the effect of BRD9 inhibition or knockdown on matrix metabolism and pyroptosis. We found that the expression of BRD9 was upregulated as IDD progressed. BRD9 inhibition or knockdown alleviated TNF-α-induced matrix degradation, reactive oxygen species (ROS) production, and pyroptosis in rat nucleus pulposus cells. Mechanistically, RNA-seq was used to investigate the mechanism of BRD9 in promoting IDD. Further investigation revealed that BRD9 regulated NOX1 expression. Inhibition of NOX1 could abrogate matrix degradation, ROS production, and pyroptosis caused by BRD9 overexpression. In vivo, the radiological and histological evaluation showed that the pharmacological inhibition of BRD9 alleviated IDD development in rat IDD model. Our results indicated that BRD9 could promote IDD via the NOX1/ROS/ NF-κB axis by inducing matrix degradation and pyroptosis. Targeting BRD9 may be a potential therapeutic strategy in treating IDD.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Materials Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Bullet Edits Limited for editing and proofreading of the article.

Funding

This work was supported by the grants from the National Natural Science Foundation of China (Nos. 81572197 and 82002776), the Natural Science Foundation of Guangdong Province (Nos. 2020A1515011538 and 2020A1515010060), the Guangdong Basic and Applied Basic Research Foundation (No. 2021A1515010345), and the Yat-sen Qihang Foundation of Sun Yat-sen memorial hospital (No. YXQH202204).

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  1. Zhihuai Deng, Yangyang Zhang, and Yuanxin Zhu contributed equally to this work.

Authors and Affiliations

  1. Department of Spine Surgery, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University, Guangzhou, China

    Zhihuai Deng, Yangyang Zhang, Yuanxin Zhu, Shuangxing Li, Zhengqi Huang, Tianyu Qin, Jiajun Wu, Chao Zhang, Dongsheng Huang & Wei Ye

  2. Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University, Guangzhou, China

    Zhihuai Deng, Yangyang Zhang, Yuanxin Zhu, Jianxiong Zhu, Shuangxing Li, Zhengqi Huang, Jiajun Wu, Chao Zhang, Weijian Chen & Wei Ye

  3. Department of Orthopedics, the Eighth Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China

    Tianyu Qin

  4. Department of Orthopedics, the Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Jianxiong Zhu & Weijian Chen

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Contributions

WY, DSH, and WJC conceived and designed the study. ZHD, YYZ, and YXZ performed experiments; JXZ and SXL prepared figures; ZQH and TYQ analyzed data; JJW and CZ helped to write and revise the manuscript. All the authors read and approved the final manuscript.

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Correspondence to Wei Ye.

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The usage of human samples was approved by the Ethics Committee of Sun Yat-Sen Memorial Hospital [No. (2022) 413]. The animal study was approved by Institutional Animal Care and Use Committee of Sun Yat-sen University (SYSU-IACUC-2022-B1211).

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Deng, Z., Zhang, Y., Zhu, Y. et al. BRD9 Inhibition Attenuates Matrix Degradation and Pyroptosis in Nucleus Pulposus by Modulating the NOX1/ROS/NF-κB axis. Inflammation 46, 1002–1021 (2023). https://doi.org/10.1007/s10753-023-01786-6

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