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Methyl Ferulic Acid Alleviates Neuropathic Pain by Inhibiting Nox4-induced Ferroptosis in Dorsal Root Ganglia Neurons in Rats

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Abstract

Neuropathic pain is a disease that has become one of the major public health problems and a global burden. Nox4-induced oxidative stress can lead to ferroptosis and neuropathic pain. Methyl ferulic acid (MFA) can inhibit the Nox4-induced oxidative stress. This study aimed to estimate whether methyl ferulic acid alleviates neuropathic pain by inhibiting the expression of Nox4 and its induction of ferroptosis. Adult male Sprague–Dawley rats were subjected to spared nerve injury (SNI) model to induce neuropathic pain. After the establishment of the model, methyl ferulic acid was given 14 days by gavage. Nox4 overexpression was induced by microinjection of the AAV-Nox4 vector. All groups measured paw mechanical withdrawal threshold (PMWT), paw thermal withdrawal latency (PTWL), and paw withdrawal cold duration (PWCD). The expression of Nox4, ACSL4, GPX4, and ROS was investigated by Western blot and immunofluorescence staining. The changes in iron content were detected by a tissue iron kit. The morphological changes in mitochondria were observed by transmission electron microscopy. In the SNI group, the paw mechanical withdrawal threshold, the paw withdrawal cold duration decreased, the paw thermal withdrawal latency did not change, the Nox4, ACSL4, ROS, and iron content increased, the GPX4 decreased, and the number of abnormal mitochondria increased. Methyl ferulic acid can increase PMWT and PWCD but does not affect PTWL. Methyl ferulic acid can inhibit Nox4 protein expression. Meanwhile, ferroptosis-related protein ACSL4 expression was decreased, GPX4 expression was increased, ROS, iron content and abnormal mitochondrial number were decreased. By overexpressing Nox4, the PMWT, PWCD, and ferroptosis of rats were more severe than those of the SNI group, but they could be reversed after treatment with methyl ferulic acid. In conclusion, methyl ferulic acid can alleviate neuropathic pain, which is related to Nox4-induced ferroptosis.

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Data availability

The data of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank American Journal Experts (www.aje.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This study was supported by the National Natural Science Foundation of China (No. 82160235) and the Scientific Research Project of Shihezi University (No. ZZZC202068A).

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  1. Tielong Liu and Ruixue Wang are authors have contributed equally to this work

Authors and Affiliations

  1. Department of Anesthesiology, First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi, China

    Tielong Liu, Ruixue Wang, Wenqiang Qi, Lei Jia, Jieting Yin, Yujia Zhao, Zhigang Dai & Jiangwen Yin

  2. NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases, First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi, China

    Tielong Liu, Ruixue Wang, Wenqiang Qi, Lei Jia, Jieting Yin, Yujia Zhao, Zhigang Dai & Jiangwen Yin

  3. Department of Physiology, School of Medicine, Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University, Shihezi, China

    Ketao Ma & Junqiang Si

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Contributions

J-W Y, R-X W, and Z-G D contributed to the study design and revised the manuscript. T-L L, W-Q Q, and L J conducted the experiment and collected and analyzed the data. K-T M, J-Q S, J-T Y, and Y-J Z provided assistance in performing the experiments. T-L L and R-X W wrote the manuscript. All authors gave final approval of the manuscript.

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Correspondence to Jiangwen Yin.

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Liu, T., Wang, R., Qi, W. et al. Methyl Ferulic Acid Alleviates Neuropathic Pain by Inhibiting Nox4-induced Ferroptosis in Dorsal Root Ganglia Neurons in Rats. Mol Neurobiol 60, 3175–3189 (2023). https://doi.org/10.1007/s12035-023-03270-6

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