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Htr2b Promotes M1 Microglia Polarization and Neuroinflammation after Spinal Cord Injury via Inhibition of Neuregulin-1/ErbB Signaling

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Abstract

The secondary injury of spinal cord injury (SCI) is dominated by neuroinflammation, which was caused by microglia M1 polarization. This study aimed to investigate the role and mechanism of Htr2b on neuroinflammation of SCI. The BV2 and HMC3 microglia were treated with lipopolysaccharide (LPS) or interferon (IFN)-γ to simulate in vitro models of SCI. Sprague–Dawley rats were subjected to the T10 laminectomy to induce animal model of SCI. Htr2b mRNA expression was measured by qRT-PCR. The expression of Htr2b and Iba-1 was detected by western blot and immunofluorescence. The expression of inflammatory cytokines in vitro and in vivo was also measured. Kyoto Encyclopedia of Genes and Genomes (KEGG) was employed to analyze Htr2b-regulated signaling pathways. Rat behavior was analyzed by the Basso, Beattie, and Bresnahan (BBB) and inclined plane test. Rat dorsal horn tissues were stained by hematoxylin–eosin (H&E) and Nissl to measure neuron loss. Htr2b was highly expressed in LPS- and IFN-γ-treated microglia and SCI rats. SCI modeling promoted M1 microglia polarization and increased levels of inflammatory cytokines. Inhibition of Htr2b by Htr2b shRNA or RS-127445 reduced the expression of Htr2b, Iba-1, and iNOS and suppressed cytokine levels. KEGG showed that Htr2b inhibited ErbB signaling pathway. Inhibition of Htr2b increased protein expression of neuregulin-1 (Nrg-1) and p-ErbB4. Inhibition of the ErbB signaling pathway markedly reversed the effect of Htr2b shRNA on M1 microglia polarization and inflammatory cytokines. Htr2b promotes M1 microglia polarization and neuroinflammation after SCI by inhibiting Nrg-1/ErbB signaling pathway.

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Data availability

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

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Authors and Affiliations

  1. Department of Orthopaedics, Qilu Hospital of Shandong University, No. 107, Wenhuaxi Road, Lixia District, 250012, Jinan, Shandong, People’s Republic of China

    Wenhao Chen, Xianlei Gao, Wanliang Yang, Xun Xiao, Xin Pan & Hao Li

  2. Cheeloo College of Medicine, Shandong University, 250012, Jinan, Shandong, People’s Republic of China

    Wenhao Chen, Wanliang Yang & Xun Xiao

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  1. Wenhao Chen
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  2. Xianlei Gao
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  5. Xin Pan
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  6. Hao Li
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Contributions

Conception and design: WH C and XL G; Perform research: WL Y and X X; Data analysis and interpretation: X P and H L; Manuscript writing: All authors; Final approval of manuscript: All authors.

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Correspondence to Hao Li.

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The experimental protocol of our study was performed in accordance with the Guide for the Care and Use of Laboratory Animals and approved by the Qilu Hospital, Cheeloo College of Medicine, Shandong University.

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Wenhao Chen and Xianlei Gao are Co-first author.

Highlights

1. Htr2b is highly expressed in LPS- and IFN-γ-treated microglia and SCI rats;.

2. Inhibition of Htr2b represses M1 microglia polarization and inflammation;.

3. Htr2b inhibits the Nrg-1/ErbB signaling.

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Chen, W., Gao, X., Yang, W. et al. Htr2b Promotes M1 Microglia Polarization and Neuroinflammation after Spinal Cord Injury via Inhibition of Neuregulin-1/ErbB Signaling. Mol Neurobiol 61, 1643–1654 (2024). https://doi.org/10.1007/s12035-023-03656-6

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