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Identification of the CCL2 PI3K/Akt axis involved in autophagy and apoptosis after spinal cord injury

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Abstract

Spinal cord injury (SCI) is a devastating neurological disease with no cure that usually results in irreversible loss of sensory and voluntary motor functions below the injury site. We conducted an in-depth bioinformatics analysis combining the gene expression omnibus spinal cord injury database and the autophagy database and found that the expression of the autophagy gene CCL2 was significantly upregulated and the PI3K/Akt/mTOR signaling pathway was activated after SCI. The results of the bioinformatics analysis were verified by constructing animal and cellular models of SCI. We then used small interfering RNA to inhibit the expression of CCL2 and PI3K to inhibit and activate the PI3K/Akt/mTOR signaling pathway; western blot, immunofluorescence, monodansylcadaverine, and cell flow techniques were used to detect the expression of key proteins involved in downstream autophagy and apoptosis. We found that when PI3K inhibitors were activated, apoptosis decreased, the levels of autophagy-positive proteins LC3-I/LC3-II and Bcl-1 increased, the levels of autophagy-negative protein P62 decreased, the levels of pro-apoptotic proteins Bax and caspase-3 decreased, the levels of the apoptosis-inhibiting protein Bcl-2 increased. In contrast, when a PI3K activator was used, autophagy was inhibited, and apoptosis was increased. This study revealed the effect of CCL2 on autophagy and apoptosis after SCI through the PI3K/Akt/mTOR signaling pathway. By blocking the expression of the autophagy-related gene CCL2, the autophagic protective response can be activated, and apoptosis can be inhibited, which may be a promising strategy for the treatment of SCI.

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Funding

This study was supported by the National Natural Science Foundation of China, No. 81871785.

And Hefei Second People’s Hospital Doctoral Special Fund Project Grant No. 2022bszx08.

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Author notes

  1. Sheng Fang, Hao Tang, Ming-Zhi Li, are co-first authors who contributed equally to this work.

Authors and Affiliations

  1. Department of Orthopedics, Hefei hospital Affiliated to Anhui Medical University, Guang De Road, 230011, Hefei, Anhui Province, People’s Republic of China

    Sheng Fang, Jian-Jun Chu & Qi-Yu Jia

  2. Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, #218 Jixi Road, 230022, Hefei, China

    Hao Tang, Ming-Zhi Li & Zong-Sheng Yin

  3. Department of Orthopedics, the Linquan county people’s Hospital Affiliated to Anhui Medical University, 109 Tongyang Road, 236400, Fuyang, Anhui Province, People’s Republic of China

    Ming-Zhi Li

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  1. Sheng Fang
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  3. Ming-Zhi Li
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Contributions

Sheng Fang: Conceived the original idea and designed the outlines of the study. Hao Tang: Helped do some experiments, Ming-zhi Li, Jian-Jun Chu Helped collect, organize the data. Zong-Sheng Yin, Qi-yu Jia: Aided in revising the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Zong-Sheng Yin or Qi-Yu Jia.

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The authors declare that they have no conflict of interest with the Gene Expression Omnibus (GEO) database used in this study.

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This study was approved by the Ethics Committee of Anhui Medical University (20180402).

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Fang, S., Tang, H., Li, MZ. et al. Identification of the CCL2 PI3K/Akt axis involved in autophagy and apoptosis after spinal cord injury. Metab Brain Dis 38, 1335–1349 (2023). https://doi.org/10.1007/s11011-023-01181-y

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