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Updated: Oct 8, 2025
Author Spotlight: Enhancing Understanding and Treatment Strategies with the NEC-on-a-Chip Model
Published on: July 28, 2023
miR-200a-3p improves neonatal necrotizing enterocolitis by regulating RIPK1
Yulu Liu1, Zhansheng Wang1, Hua Huang2
1Department of Neonatal Intensive Care Unit, The First People's Hospital of Shangqiu Shangqiu 476100, Henan Province, China.
Background:
Necrotizing enterocolitis (NEC) is an acquired disease, which mainly occurs in premature infants or sick newborns. microRNA (miR), as a common non-coding RNA in recent years, is found in many diseases. In this research, miR usefulin NEC is analyzed by GEO.
Method:
The differentially expressed miRs in NEC were screened by analyzing GSE68054, and miR-200a-3p in IEC-6 cells induced by lipopolysaccharide (LPS) and serum of NEC children were detected by qRT-PCR. The role of miR-200a-3p in LPS-induced IEC-6 cells was tested using CCK-8, PI dyeing, and inflammatory cytokine detection. The direct downstream molecules of miR-200a-3p were identified using TargetScanHuman and verified by luciferase reporter gene assay. The mechanism of action was explored using western blot.
Results:
miR-200a-3p in IEC-6 treated with NEC and LPS was significantly decreased. In vitro experiments revealed that miR-200a-3p mimetic could inhibit IL-6 and TNF-α in IEC-6 cells induced by LPS and reduce the positive rate of PI. In addition, it was determined that receptor-interacting protein kinase 1 (RIPK1) was a downstream molecule of miR-200a-3p, and overexpression of RIPK1 could aggravate LPS-induced IEC-6 injury, while miR-200a-3p mimics could alleviate the overexpression of RIPK1. miR-200a-3p mimics inhibited the elevation of necrosis-related molecules and the interaction between RIPK1 and RIPK3 in LPS-induced IEC-6 cells.
Conclusion:
miR-200a-3p can protect intestinal epithelial cells from LPS injury by inhibiting inflammation and necrosis mediated by RIPK1, which provides a possible target for NEC.
Insights
MicroRNA-200a-3p protects intestinal cells from necrotizing enterocolitis (NEC) by reducing inflammation and necrosis. This microRNA (miRNA) targets RIPK1, offering a potential therapeutic avenue for NEC treatment.
Area of Science:
- Gastroenterology
- Molecular Biology
- Neonatology
Background:
- Necrotizing enterocolitis (NEC) is a severe acquired intestinal disease primarily affecting premature infants.
- MicroRNAs (miRNAs) are non-coding RNAs implicated in various disease pathologies.
- This study investigates the role of specific miRNAs in the context of NEC.
Purpose of the Study:
- To identify differentially expressed microRNAs in necrotizing enterocolitis (NEC).
- To elucidate the functional role and mechanism of miR-200a-3p in intestinal epithelial cells under NEC-related stress.
- To explore miR-200a-3p as a potential therapeutic target for NEC.
Main Methods:
- Differential expression analysis of miRNAs using GEO dataset GSE68054.
- Quantitative real-time PCR (qRT-PCR) to measure miR-200a-3p levels in lipopolysaccharide (LPS)-induced IEC-6 cells and NEC patient serum.
- In vitro assays (CCK-8, PI staining, cytokine detection) to assess cell viability, death, and inflammation.
- Bioinformatic prediction (TargetScanHuman) and luciferase reporter assays to identify miR-200a-3p targets.
- Western blot analysis to investigate molecular mechanisms, including RIPK1 interactions.
Main Results:
- miR-200a-3p expression was significantly decreased in LPS-treated IEC-6 cells and NEC contexts.
- miR-200a-3p mimics inhibited LPS-induced inflammation (IL-6, TNF-α) and cell death (PI positive rate) in IEC-6 cells.
- Receptor-interacting protein kinase 1 (RIPK1) was identified as a direct downstream target of miR-200a-3p.
- Overexpression of RIPK1 exacerbated LPS-induced cell injury, while miR-200a-3p mimics alleviated RIPK1 levels and inhibited RIPK1-RIPK3 interactions, reducing necrosis.
Conclusions:
- miR-200a-3p confers protection to intestinal epithelial cells against LPS-induced injury.
- This protection is achieved by suppressing inflammation and necrosis pathways mediated by RIPK1.
- miR-200a-3p represents a promising therapeutic target for necrotizing enterocolitis.
