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MiRNA-192-5p-targeted activated leukocyte cell adhesion molecule improved inflammatory injury of neonatal necrotizing enterocolitis

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Abstract

Background

Neonatal necrotizing enterocolitis (NEC) is a common gastrointestinal emergency in neonates. MiRNA-192-5p was found associated with ulcerative colitis (UC) progression, also with aberrant expression in intestinal cancer tissue. However, the effects of miRNA-192-5p on NEC have not been reported.

Methods

Based on the bioinformatics analysis of the GEO dataset, miR-192-5p was identified as the differentially expressed miRNA in NEC, and activated leukocyte cell adhesion molecule (ALCAM) was predicted as its target. After that, in vitro, rat intestinal epithelial cell-6 (IEC-6) were stimulated with LPS to construct a cell model of NEC. IEC-6 cells were transfected with miRNA-192-5p mimics, miRNA-192-5p inhibitors, or miRNA-192-5p inhibitors + sh-ALCAM, and relevant negative control. In vivo, SD rats were treated with artificial feeding, hypoxic reoxygenation, cold stimulation, and LPS gavage to induce NEC, followed by injection of agomiR-NC or agomiRNA-192-5p. Then effects of miRNA-192-5p on NEC model IEC-6 cell viability, apoptosis, ALCAM expression, Interleukin (IL)-1β and IL-6 levels, intestinal injury, intestinal permeability were detected.

Results

MiRNA-192-5p expression was downregulated in NEC IEC-6 cells, whose overexpression increased IEC-6 cell viability. MiRNA-192-5p inhibitors increased IL-1β, IL-6 levels and promoted IEC-6 cell apoptosis. MiRNA-192-5p targeting of ALCAM decreased ALCAM expression, IL-1β, and IL-6 levels. AgomiRNA-192-5p decreased ALCAM, IL-1β, and IL-6 levels in intestinal tissue and pathological damage and increased miRNA-192-5p levels.

Conclusion

MiR-192-5p protects against intestinal injury by inhibiting ALCAM-mediated inflammation and intestinal epithelial cells, which would provide a new idea for NEC treatment.

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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.

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Funding

This work was supported by the program of Department of Health of Zhejiang Province [2022KY1012].

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Authors and Affiliations

  1. Department of Pediatric Surgery, Hangzhou Children’s Hospital, Hangzhou, 310005, Zhejiang, China

    Zhenfei Wu, You Tian, Chen Wang & Jie Zhang

  2. Department of Pediatric Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, No. 109, West Xueyuan Road, Wenzhou, 325027, Zhejiang, China

    Jinhan Lin

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Contributions

Conception and design of this research: Zhenfei Wu, Acquisition of data: You Tian, Interpretation of the data: Chen Wang, Analysis of the data: Jie Zhang, Drafting the manuscript: Zhenfei Wu, Revision of the manuscript for important intellectual content: Jinhan Lin, Approval of final version of manuscript to be published: All authors, Agreement to be accountable for all aspects of the work ensuring that questions related to the accuracy or integrity of the work are appropriately investigated and resolved: Zhenfei Wu, You Tian, Chen Wang, Jie Zhang, Jinhan Lin.

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Correspondence to Jinhan Lin.

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All animal-related procedures were adhered to the Institutional Animal Care and Use Committee. Ethics approval was obtained from the Animal Experimentation Ethics Committee of Zhejiang Eyong Pharmaceutical Research and Development Center (Certificate No. SYXK (Zhe) 2021–0033).

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Wu, Z., Tian, Y., Wang, C. et al. MiRNA-192-5p-targeted activated leukocyte cell adhesion molecule improved inflammatory injury of neonatal necrotizing enterocolitis. Pediatr Surg Int 40, 126 (2024). https://doi.org/10.1007/s00383-024-05713-0

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