Abstract
Paralytic ileus is common in patients with septic shock, causing high morbidity and mortality. Enteric neurons and enteric glial cells (EGCs) regulate intestinal motility. However, little is known about their interaction in endotoxemia. This study aimed to investigate whether reactive EGCs had harmful effects on enteric neurons and participated in intestinal motility disorder in mice during endotoxemia. Endotoxemia was induced by the intraperitoneal injection of lipopolysaccharide (LPS) in mice. Fluorocitrate (FC) was administered before LPS injection to inhibit the reactive EGCs. The effects of reactive EGCs on intestinal motility were analyzed by motility assays in vivo and colonic migrating motor complexes ex vivo. The number of enteric neurons was evaluated by immunofluorescent staining of HuCD, nNOS, and ChAT in vivo. In addition, we stimulated EGCs with IL-1β and TNF-α in vitro and cultured the primary enteric neurons in the conditioned medium, detecting the apoptosis and morphology of neurons through staining TUNEL, cleaved caspase-3 protein, and anti-β-III tubulin. Intestinal motility and peristaltic reflex were improved by inhibiting reactive EGCs in vivo. The density of the neuronal population in the colonic myenteric plexus increased significantly, while the reactive EGCs were inhibited, especially the nitrergic neurons. In vitro, the enteric neurons cultured in the conditioned medium of reactive EGCs had a considerably higher apoptotic rate, less dendritic complexity, and fewer primary neurites. Reactive enteric glial cells probably participated in paralytic ileus by damaging enteric neurons during endotoxemia. They might provide a novel therapeutic strategy for intestinal motility disorders during endotoxemia or sepsis.
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The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We want to express our gratitude to Professor Shengxi Wu and his colleagues (Department of Neurobiology, Air Force Medical University, Xi’an, Shaanxi Province, China) who shared equipment and reagents generously in getting the preliminary data.
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This work was supported by funding from the National Natural Science Foundation of China (Nos. 81774113 and 81974540), Beijing, China, and the Clinical Research Award of the First Affiliated Hospital of Xi’an Jiaotong University (No. XJTU1AF-CRF-2016–003), Xi’an, China.
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Qiang Wang and Shuang Li designed experiments and developed methodologies. Material preparation, experiments, and data collection were performed by Na Li, Hui Gao, and Jing Xu. Yuxin Zhang and Haiqing Chang analyzed and interpreted the data. The first draft of the manuscript was written by Na Li, and all authors commented on previous versions of the manuscript. Qiang Wang, Jing Xu, and Shuwen Tan revised the manuscript. All authors read and approved the final manuscript.
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Li, N., Xu, J., Gao, H. et al. Effect of Reactive EGCs on Intestinal Motility and Enteric Neurons During Endotoxemia. J Mol Neurosci 72, 1831–1845 (2022). https://doi.org/10.1007/s12031-022-02044-4
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DOI: https://doi.org/10.1007/s12031-022-02044-4