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Neuron-derived exosomes mediate sevoflurane-induced neurotoxicity in neonatal mice via transferring lncRNA Gas5 and promoting M1 polarization of microglia

Acta Pharmacologica Sinica volume 45, pages 298–311 (2024)Cite this article

Abstract

Sevoflurane exposure during rapid brain development induces neuronal apoptosis and causes memory and cognitive deficits in neonatal mice. Exosomes that transfer genetic materials including long non-coding RNAs (lncRNAs) between cells play a critical role in intercellular communication. However, the lncRNAs found in exosomes derived from neurons treated with sevoflurane and their potential role in promoting neurotoxicity remain unknown. In this study, we investigated the role of cross-talk of newborn mouse neurons with microglial cells in sevoflurane-induced neurotoxicity. Mouse hippocampal neuronal HT22 cells were exposed to sevoflurane, and then co-cultured with BV2 microglial cells. We showed that sevoflurane treatment markedly increased the expression of the lncRNA growth arrest-specific 5 (Gas5) in neuron-derived extracellular vesicles, which inhibited neuronal proliferation and induced neuronal apoptosis by promoting M1 polarization of microglia and the release of inflammatory cytokines. We further revealed that the exosomal lncRNA Gas5 significantly upregulated Foxo3 as a competitive endogenous RNA of miR-212-3p in BV2 cells, and activated the NF-κB pathway to promote M1 microglial polarization and the secretion of inflammatory cytokines, thereby exacerbating neuronal damage. In neonatal mice, intracranial injection of the exosomes derived from sevoflurane-treated neurons into the bilateral hippocampi significantly increased the proportion of M1 microglia, inhibited neuronal proliferation and promoted apoptosis, ultimately leading to neurotoxicity. Similar results were observed in vitro in BV2 cells treated with the CM from HT22 cells after sevoflurane exposure. We conclude that sevoflurane induces the transfer of lncRNA Gas5-containing exosomes from neurons, which in turn regulates the M1 polarization of microglia and contributes to neurotoxicity. Thus, modulating the expression of lncRNA Gas5 or the secretion of exosomes could be a strategy for addressing sevoflurane-induced neurotoxicity.

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Fig. 1: Neuron-derived exosomes after sevoflurane treatment increase neuron apoptosis by promoting the polarization of M1 microglia.
Fig. 2: Increased lncRNA Gas5 aggravates the polarization of M1 microglia and the secretion of inflammatory cytokines.
Fig. 3: Gas5 functions as a ceRNA for miR-212-3p in BV2 cells.
Fig. 4: Binding between miR-212-3p and Foxo3 and its effects on the secretion of inflammatory cytokines.
Fig. 5: lncRNA Gas5 promotes the polarization of M1 microglia, the secretion of inflammatory cytokines, and neuronal apoptosis by regulating Foxo3.
Fig. 6: lncRNA Gas5 activates NF-κB p65 subunit nuclear importation.
Fig. 7: lncRNA Gas5 promotes the polarization of M1 microglia and learning and memory impairments in mice.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (82271287), the Exploration Project of Zhejiang Natural Science Foundation (LY21H090006), the Zhejiang Health Science and Technology Planning Project (2021KY768), the Bureau of Chinese Medicine, Zhejiang, China (2018ZB065). This work was partly supported by grants from the Nanhu Brain-computer Interface Institute (010902001).

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Authors and Affiliations

  1. Department of Anesthesia, Women’s Hospital, Zhejiang University School of Medicine, Hangzhou, 310006, China

    Li-li Xu, Jia-qian Xie & Xin-zhong Chen

  2. Department of Anesthesia, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China

    Jian-jun Shen

  3. Nanhu Brain-computer Interface Institute, Hangzhou, 311100, China

    Mei-dan Ying

  4. Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China

    Mei-dan Ying

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Contributions

LLX, MDY, and XZC conceived and designed the experiments. LLX and JQX performed the data analysis work and wrote the paper. JJS collected and processed the data; XZC revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Li-li Xu, Mei-dan Ying or Xin-zhong Chen.

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Xu, Ll., Xie, Jq., Shen, Jj. et al. Neuron-derived exosomes mediate sevoflurane-induced neurotoxicity in neonatal mice via transferring lncRNA Gas5 and promoting M1 polarization of microglia. Acta Pharmacol Sin 45, 298–311 (2024). https://doi.org/10.1038/s41401-023-01173-9

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