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Low-dose Lipopolysaccharide Alleviates Spinal Cord Injury-induced Neuronal Inflammation by Inhibiting microRNA-429-mediated Suppression of PI3K/AKT/Nrf2 Signaling

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Abstract

This study investigated the impact of low-dose lipopolysaccharide (LPS) on spinal cord injury (SCI) and the potential molecular mechanism. Rats were randomly assigned to four groups: Sham, SCI, SCI + LPS, and SCI + LPS + agomir. Allen’s weight-drop method was used to establish an in vivo SCI model. The Basso Bcattie Bresnahan rating scale was employed to monitor locomotor function. An in vitro SCI model was constructed by subjecting PC12 cells to oxygen and glucose deprivation/ reoxygenation (OGD/R). Enzyme-linked immunosorbent assay (ELISA) was applied for the determination interleukin (IL)-1β and IL-6. The dual luciferase reporter assay was used to validate the targeting of microRNA (miR)-429 with PI3K. Immunohistochemical staining was used to assess the expression of PI3K, phosphorylated AKT and Nrf2 proteins. The Nrf2-downstream anti-oxidative stress proteins, OH-1 and NQO1, were detected by western blot assay. MiR-429 expression was detected by fluorescence in situ hybridization and real-time quantitative reverse transcription PCR. In vitro, low-dose LPS decreased miR-429 expression, activated PI3K/AKT/Nrf2, inhibited oxidative stress and inflammation, and attenuated SCI. MiR-429 was found to target and negatively regulate PI3K. Inhibition of miR-429 suppressed low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. In vivo, miR-429 was detectable in neurons. Inhibition of miR-429 blocked low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. Overall, low-dose LPS was found to alleviate SCI-induced neuronal oxidative stress and inflammatory response by down-regulating miR-429 to activate the PI3K/AKT/Nrf2 pathway.

Graphical Abstract

Highlights

Low-dose LPS reduces the inflammatory response and alleviates SCI.

Low-dose LPS decreases miR-429 levels and activates PI3K/AKT/Nrf2 signaling in SCI.

MiR-429 targets PI3K in SCI.

Inhibition of miR-429 activates PI3K/AKT/Nrf2 signaling in SCI.

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Data Availability

Data will be made available on reasonable request.

Code Availability

Not applicable.

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Acknowledgements

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Funding

This study was supported by the National Natural Science Foundation of China (Grant No.: 82260257), Key Research Project of Yunnan Provincial Science and Technology (Grant No.: 202102AA310042; 202001AS070028), Key project of Yunnan clinical medicine research center (Grant No.: 2022YJZX-GK02; 2022YJZX-GK23).

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Author notes

  1. Weichao Li and Tao Tang contributed equally to this work and are co-first authors.

Authors and Affiliations

  1. Department of Orthopedic Surgery, The First People’s Hospital of Yunnan province, Affiliated Hospital of Kunming University of Science and Technology, Kunming, 650032, China

    Weichao Li, Shaoping Yao & Tiannan Zou

  2. Faculty of Medical Science, Kunming University of Science and Technology, Kunming, 650500, China

    Weichao Li, Tao Tang, Shixiao Zhong & Qianbo Fan

  3. Yunnan Key Laboratory of Digital Orthopaedics, Kunming, 650032, China

    Weichao Li, Shaoping Yao & Tiannan Zou

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  1. Weichao Li
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Contributions

Weichao Li, Tao Tang, and Tiannan Zou performed the experiment; Shaoping Yao, Qianbo Fan, and Shixiao Zhong contributed significantly to analysis and manuscript preparation; Weichao Li, Tao Tang, and Tiannan Zou performed the data analyses and wrote the manuscript; Weichao Li, Tao Tang, and Tiannan Zou helped perform the analysis with constructive discussions. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Tiannan Zou.

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All animals were kept in a pathogen-free environment and fed ad lib. The procedures for care and use of animals were approved by the Ethics Committee of the First People’s Hospital of Yunnan province and all applicable institutional and governmental regulations concerning the ethical use of animals were followed.

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Li, W., Tang, T., Yao, S. et al. Low-dose Lipopolysaccharide Alleviates Spinal Cord Injury-induced Neuronal Inflammation by Inhibiting microRNA-429-mediated Suppression of PI3K/AKT/Nrf2 Signaling. Mol Neurobiol 61, 294–307 (2024). https://doi.org/10.1007/s12035-023-03483-9

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