Related Experiment Video
Updated: Sep 11, 2025
Modeling Encephalopathy of Prematurity Using Prenatal Hypoxia-ischemia with Intra-amniotic Lipopolysaccharide in Rats
Published on: November 20, 2015
Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm
Tyler C Hillman1,2, Braeden Jacobson2, Kiara Piaggio Hurtado De Medoza3
1Lawrence D. Longo, MD Center for Perinatal Biology, Department of Basic Science, Loma Linda University, Loma Linda, CA, United States.
Insights
A new murine model for preterm hypoxic-ischemic encephalopathy (pHIE) effectively replicates human brain injury. This model shows increased inflammation and neuronal damage, offering a cost-effective research alternative for studying pHIE.
Area of Science:
- Neuroscience
- Developmental Biology
- Pathology
Background:
- Preterm hypoxic-ischemic encephalopathy (pHIE) is a significant cause of neurological disorders in newborns.
- Current interventions for pHIE are ineffective, highlighting the need for better models to study the condition.
- Existing animal models are often costly or do not fully capture the complexity of pHIE.
Purpose of the Study:
- To develop and characterize a cost-effective murine model for preterm hypoxic-ischemic encephalopathy (pHIE).
- To investigate the inflammatory and neuronal damage associated with pHIE in this new model.
- To provide a valuable research tool for understanding and potentially treating pHIE.
Main Methods:
- Pregnant mice were exposed to LPS to induce inflammation, followed by hypoxia in pups.
- Immunohistochemistry and unbiased stereology were used to quantify Caspase-9 expression.
- Machine learning-based image analysis assessed MAP2 expression, and RT-qPCR/ELISA measured cytokine levels.
Main Results:
- pHIE pups exhibited significantly elevated Caspase-9 expression in the cortex.
- MAP2 expression, a marker of neuronal health, was significantly decreased in pHIE pups.
- Key inflammatory cytokines (IL-1β, IL-10) were significantly upregulated in the pHIE model.
Conclusions:
- The developed murine model successfully replicates key aspects of human pHIE, including cortical damage and inflammation.
- This model serves as a viable and cost-effective alternative for studying HIE.
- Further research using this model can advance understanding and therapeutic strategies for pHIE.
Introduction:
Preterm hypoxic-ischemic encephalopathy (pHIE) is a complex brain injury that contributes to chronic neural inflammation and neurological disorders. The signs and symptoms of in utero pHIE can often be overlooked, untreated or lumped into more generic conditions such as encephalopathy of prematurity (EOP). Clinical interventions like hypothermia and erythropoietin do not improve pHIE. We characterized a murine model for pHIE, which includes hypoxia and maternal factors as a cost-effective alternative to large animal models of HIE.
Methods:
We injected pregnant mouse dams with LPS to stimulate an inflammatory response on embryonic days 15-16 (E15-E16), and whole cage hypoxia exposures occurred from postnatal days 3 to 9. To quantify the development of inflammation in the pHIE model, we used immunohistochemistry to stain for Caspase-9 in the cortex (20 μm per slice) and then counted Caspase-9 positive cells using unbiased stereology. We stained brain tissue with MAP2 to quantify neuronal intermediate filament expression and staining using a machine-learning based image analysis approach. We quantified cytokines (IL-1β, IL-6, IL-10, IL-18 and TNF-α) using RT-qPCR and (IL-18) ELISA to characterize differential expression in all treatment groups. The pHIE animals were compared with controls (LPS-Normoxia, Saline-Hypoxia, Saline-Normoxia, and Naïve) and with a model of only hypoxia (10% O2) exposure in mouse pups.
Results:
The pHIE pups showed significantly higher expression of Caspase-9 throughout the cortex compared to Naïve pup brains (p < 0.05). MAP2 expression was significantly decreased (p < 0.05) between 1.5-6.0 mm of the brain compared to Saline-Hypoxia and Naïve animals. Both IL-1β and IL-10 expression in LPS-Hypoxia animals was significantly higher (p < 0.05) than in Saline-Hypoxia and Naive animals. TNF-α expression was not significantly different between LPS-Hypoxia and Saline-Hypoxia animals. However, both showed significantly different transcription, compared to Naive animals.
Discussion:
The model we describe here shows cortical damage similar to that seen in human HIE.
More Related Videos
08:50A Murine Model of Fetal Exposure to Maternal Inflammation to Study the Effects of Acute Chorioamnionitis on Newborn Intestinal Development
Published on: June 24, 2020
09:12Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014