Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy
IntroductionPreterm 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...
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Frontiers Media S.A.
2025-07-01
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| Series: | Frontiers in Systems Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fsysb.2025.1517712/full |
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| author | Tyler C. Hillman Tyler C. Hillman Braeden Jacobson Kiara Piaggio Hurtado De Medoza Marlene Lopez Nicholas Iwakoshi Christopher G. Wilson Christopher G. Wilson Christopher G. Wilson |
| author_facet | Tyler C. Hillman Tyler C. Hillman Braeden Jacobson Kiara Piaggio Hurtado De Medoza Marlene Lopez Nicholas Iwakoshi Christopher G. Wilson Christopher G. Wilson Christopher G. Wilson |
| author_sort | Tyler C. Hillman |
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| description | IntroductionPreterm 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.MethodsWe 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.ResultsThe 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.DiscussionThe model we describe here shows cortical damage similar to that seen in human HIE. |
| format | Article |
| id | doaj-art-dcb22274b2914dd5b5bcbe9e4724dd5b |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-dcb22274b2914dd5b5bcbe9e4724dd5b2025-08-20T03:29:57ZengFrontiers Media S.A.Frontiers in Systems Biology2674-07022025-07-01510.3389/fsysb.2025.15177121517712Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathyTyler C. Hillman0Tyler C. Hillman1Braeden Jacobson2Kiara Piaggio Hurtado De Medoza3Marlene Lopez4Nicholas Iwakoshi5Christopher G. Wilson6Christopher G. Wilson7Christopher G. Wilson8Lawrence D. Longo, MD Center for Perinatal Biology, Department of Basic Science, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesWhittier College, Department of Biology, Whittier, CA, United StatesLawrence D. Longo, MD Center for Perinatal Biology, Department of Basic Science, Loma Linda University, Loma Linda, CA, United StatesLawrence D. Longo, MD Center for Perinatal Biology, Department of Basic Science, Loma Linda University, Loma Linda, CA, United StatesLawrence D. Longo, MD Center for Perinatal Biology, Department of Basic Science, Loma Linda University, Loma Linda, CA, United StatesSchool of Medicine, Loma Linda University, Loma Linda, CA, United StatesDepartment of Pediatrics, Loma Linda University Medical Center, Loma Linda, CA, United StatesIntroductionPreterm 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.MethodsWe 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.ResultsThe 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.DiscussionThe model we describe here shows cortical damage similar to that seen in human HIE.https://www.frontiersin.org/articles/10.3389/fsysb.2025.1517712/fullhypoxiaischemiapretermneonatematernal inflammationfetal inflammation |
| spellingShingle | Tyler C. Hillman Tyler C. Hillman Braeden Jacobson Kiara Piaggio Hurtado De Medoza Marlene Lopez Nicholas Iwakoshi Christopher G. Wilson Christopher G. Wilson Christopher G. Wilson Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy Frontiers in Systems Biology hypoxia ischemia preterm neonate maternal inflammation fetal inflammation |
| title | Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy |
| title_full | Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy |
| title_fullStr | Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy |
| title_full_unstemmed | Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy |
| title_short | Inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic-ischemic encephalopathy |
| title_sort | inflammation mediated brain damage and cytokine expression in a maternally derived murine model for preterm hypoxic ischemic encephalopathy |
| topic | hypoxia ischemia preterm neonate maternal inflammation fetal inflammation |
| url | https://www.frontiersin.org/articles/10.3389/fsysb.2025.1517712/full |
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