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MiR-26b-3p Promotes Intestinal Motility Disorder by Targeting FZD10 to Inhibit GSK3β/β-Catenin Signaling and Induce Enteric Glial Cell Apoptosis

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Abstract

Enteric glial cells (EGCs) are the major component of the enteric nervous system and affect the pathophysiological process of intestinal motility dysfunction. MicroRNAs (miRNAs) play an important role in regulating gastrointestinal homeostasis. However, the mechanism of miRNA-mediated regulation of EGCs in intestinal dysmotility remains unclear. In this study, we investigated the effect of EGC apoptosis on intestinal dysmotility, and the effect of miR-26b-3p on EGC proliferation and apoptosis in vivo and in vitro. A loperamide hydrochloride (Lop)–induced constipated mouse model and an in vitro culture system of rat EGCs were established. The transcriptome was used to predict the differentially expressed gene miR-26b-3p and the target gene Frizzled 10 (FZD10), and their targeting binding relationship was verified by luciferase. EGCs were transfected with miR-26b-3p mimic or antagomir, and the FZD10 expression was down-regulated by siRNA. Immunofluorescence and flow cytometry were used to detect EGC apoptosis. MiR-26b-3p and FZD10 expressions were examined using quantitative real-time PCR (qRT-PCR). The CCK-8 assay was used to detect EGC proliferation. The protein levels were detected by Western blotting and enzyme-linked immunosorbent assay (ELISA). The results showed that miR-26b-3p was up-regulated in the Lop group, whereas FZD10 was down-regulated, and EGC apoptosis was increased in the colon of intestinal dysmotility mice. FZD10 down-regulation and miR-26b-3p mimic significantly increased glycogen synthase kinase-3β phosphorylation (p-GSK3β) levels, decreased β-catenin expression, and promoted EGC apoptosis. MiR-26b-3p antagomir alleviated intestinal dysmotility, promoted EGC increased activity of EGCs, and reduced EGC apoptosis in vivo. In conclusion, this study indicated that miR-26b-3p promotes intestinal motility disorders by targeting FZD10 to block GSK3β/β-catenin signaling and induces apoptosis in EGCs. Our results provide a new research target for the treatment and intervention of intestinal dysmotility.

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Data Availability

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

Abbreviations

BDNF:

Brain-derived neurotrophic factor

DEMs:

Differentially expressed miRNAs

EGCs:

Enteric glial cells

ELISA:

Enzyme-linked immunosorbent assay

FZD10:

Frizzled 10

GSK3:

Glycogen synthase kinase 3

GDNF:

Glial cell–derived neurotrophic factor

GSNO:

S-Nitrosoglutathione

GFAP:

Glial fibrillary acidic protein

Lop:

Loperamide hydrochloride

miRNAs:

MicroRNAs

NGF:

Nerve growth factor

p-GSK3β:

Glycogen synthase kinase-3β phosphorylation

qRT-PCR:

Quantitative real-time PCR

S100B:

S100B protein

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Funding

This work was supported by the Youth Program of National Natural Science Foundation of China (No. 82004173), the National Natural Science Foundation of China (No. 82074429), the Science and technology Research Special project of Sichuan provincial Administration of Traditional Chinese Medicine (No. 2020JC0063), and the Science and Technology Project of Luzhou City (2022-SYF-99).

Author information

Authors and Affiliations

  1. Hospital of Chengdu University of TCM, Chengdu, China

    Yu Zhan

  2. Anorectal Department, Affiliated Hospital of Integrative Chinese Medicine and Western Medicine of Chengdu University of TCM, Chengdu, China

    Yu Zhan

  3. Chengdu First People’s Hospital, Chengdu, China

    Yu Zhan

  4. Department of Traditional Chinese Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, China

    Yong Wen

  5. Anorectal Department, People’s Hospital of Deyang City, Deyang, China

    Fan Zheng

  6. Universiti Sains Malaysia, George Town, Pulau Pinang, Malaysia

    Fan Zheng

  7. The Third People’s Hospital of Chengdu, Chengdu, China

    Li-juan Du

  8. Southwest Jiaotong University College of Medicine, Chengdu, China

    Li-juan Du

  9. Department of Integrated Traditional and Western Medicine Anorectal, Affiliated Hospital of North Sichuan Medical College, Nanchong, China

    Tai-yu Chen & Xue-gui Tang

  10. Anorectal Department, Luzhou People’s Hospital, Luzhou, China

    Xu-long Shen

  11. Chengdu University of Traditional Chinese Medicine, Chengdu, China

    Rong Wu

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Contributions

All authors have contributed significantly to this work. YZ and YW contributed equally to this work. YZ, YW, and XGT conceived and designed the experiments; FZ, LJD, and TYC performed most of the experiments; LJD and TYC contributed to data collection; XLS and RW analyzed the data; YZ and YW wrote the manuscript; XGT revised and edited the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Xue-gui Tang.

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Ethics Approval

The procedures in this experiment were performed in accordance with protocols approved by the Animal Ethics Committee of West China Hospital of Sichuan University (Approval No. 20221203006).

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The authors declare no competing interests.

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Yu Zhan and Yong Wen contributed equally to this work.

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Zhan, Y., Wen, Y., Zheng, F. et al. MiR-26b-3p Promotes Intestinal Motility Disorder by Targeting FZD10 to Inhibit GSK3β/β-Catenin Signaling and Induce Enteric Glial Cell Apoptosis. Mol Neurobiol 61, 1543–1561 (2024). https://doi.org/10.1007/s12035-023-03600-8

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