Novel Therapeutic Targeting of CCL3-CCR4 Axis Mediated Apoptotic Intesitnal Injury in Necrotizing Enterocolitis
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Updated: Sep 24, 2025
A Novel Human Epithelial Enteroid Model of Necrotizing Enterocolitis
Published on: April 10, 2019
Xi Yuan1, Zihan Xiong1, Wei Liu2
1Department of Clinical Laboratory, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Children's Hospital of Chongqing Medical University, Chongqing, China.
Background:
Necrotizing enterocolitis (NEC) is the leading cause of neonatal gastrointestinal-related death, while the etiology and pathogenesis are poorly understood.
Methods:
The levels of CCL3 in intestinal tissue from modeling mice and patients were measured and analyzed. HE staining, TUNEL, Annexin and FCM were used to assess pathological changes and apoptosis in intestinal tissue and epithelial cells. CCL3, CCR4, cytokines, tight junction protein ZO-1, apoptosis-related genes and ERK1/2-NF-κB signaling pathway were detected by ELISA, Q-PCR, Western blotting and immunofluorescence.
Results:
CCL3 levels in the intestinal tissue significantly elevated in patients with NEC and mouse models. Blockade of CCL3 significantly alleviated NEC-related intestinal tissue damage, while administration of recombinant CCL3 aggravated intestinal injury by exacerbating intestinal epithelial cell apoptosis in NEC mice. Importantly, CCR4 blockade reversed CCL3-mediated damage to intestinal tissue and intestinal epithelial cell apoptosis both in vivo and in vitro. Further mechanistic studies showed that CCL3 regulated apoptosis-related BAX/BCL-2 expression through the activation of the ERK1/2 and NF-κB pathways, which could be reversed by anti-CCR4 treatment. Furthermore, ERK1/2 inhibition reduced CCL3-mediated phosphorylation of NF-κB in IEC-6 cells, while inhibition of NF-κB had no obvious effect on ERK1/2 phosphorylation. As expected, inhibition of NF-κB regulated BAX/BCL-2 expression and alleviated CCL3-induced epithelial cell apoptosis. These results indicate that high expression of CCL3 in NEC lesions promotes intestinal epithelial apoptosis through the CCL3-CCR4-ERK1/2-NFκB-BAX/BCL2 signalling axis, thereby exacerbating NEC-related intestinal injury.
Conclusions:
Our study represents an important conceptual advance that CCL3 may be one of the key culprits of intestinal tissue damage in NEC patients, and blocking either CCL3, CCR4, or NF-κB may represent a novel effective immunotherapy for NEC.
Insights
Necrotizing enterocolitis (NEC) involves elevated CCL3 levels, which promote intestinal cell apoptosis via the CCL3-CCR4-ERK1/2-NF-κB pathway. Blocking these targets may offer new immunotherapy for NEC.
Area of Science:
- Gastroenterology
- Immunology
- Neonatal Research
Background:
- Necrotizing enterocolitis (NEC) is a primary cause of neonatal gastrointestinal mortality.
- The precise etiology and pathogenesis of NEC remain incompletely understood.
Purpose of the Study:
- To investigate the role of CCL3 in NEC pathogenesis.
- To elucidate the signaling pathways involved in CCL3-mediated intestinal injury in NEC.
Main Methods:
- Analysis of CCL3 levels in human and murine NEC intestinal tissues.
- Assessment of pathological changes and apoptosis using HE staining, TUNEL, Annexin, and FCM.
- Detection of signaling molecules (CCL3, CCR4, ERK1/2, NF-κB) and apoptosis markers (BAX/BCL-2) via ELISA, Q-PCR, Western blotting, and immunofluorescence.
Main Results:
- Elevated CCL3 levels were observed in NEC patients and mouse models.
- CCL3 blockade ameliorated NEC-induced intestinal damage and apoptosis; CCL3 administration aggravated injury.
- CCR4 blockade reversed CCL3-mediated damage and apoptosis, highlighting the CCL3-CCR4 axis.
- CCL3 induced apoptosis via the ERK1/2-NF-κB pathway, regulating BAX/BCL-2 expression.
Conclusions:
- CCL3 is a key factor in NEC-related intestinal tissue damage.
- The identified CCL3-CCR4-ERK1/2-NF-κB-BAX/BCL2 signaling axis is crucial in NEC pathogenesis.
- Targeting CCL3, CCR4, or NF-κB presents a potential novel immunotherapy for NEC.
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