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Updated: Oct 7, 2025
Early Pathological and Magnetic Resonance Detection of Cerebral Injury Using a Rat Model of Neonatal Hypoxic Ischemic Encephalopathy
Published on: October 28, 2022
Src Family Kinases Inhibition Ameliorates Hypoxic-Ischemic Brain Injury in Immature Rats
Han Qiu1, Tianyang Qian1, Tong Wu1
1National Health Commission Key Laboratory of Neonatal Diseases, Department of Neonatology, Children's Hospital of Fudan University, Shanghai, China.
Abstract:
Hypoxic-ischemic (HI) injury is one of the initial factors contributing to neonatal brain injury. Src family kinases (SFKs) are considered to act as molecular hubs for N-methyl-d-aspartate receptor (NMDAR) regulation and participate in the HI injury process. The objectives of this study were to evaluate the levels of phospho-Src (p-Src), the relationship between NMDARs and SFKs, and the effects of SFK inhibition on an immature rat HI brain injury model. The model was induced in 3-day-old Sprague-Dawley rats using the Rice-Vannucci model operation. The level of p-Src was evaluated using Western blotting. The association of NMDARs with SFKs was detected using Western blotting and coimmunoprecipitation. After intraperitoneal injection of 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo [3,4-d] pyrimidine (PP2), an SFK-selective inhibitor, neuropathological changes were observed by performing H&E and immunofluorescence staining, and the neurological functions were assessed using the following behavioral tests: modified neurological severity score, open field test, and Morris water maze test. The levels of p-Src first decreased at 0 h after injury, increased at 2 h after injury, and continuously decreased from 6 h to 3 days. Along with the increased p-Src levels observed at 2 h after injury, the phosphorylation of NMDAR subunit NR2B at tyrosine 1472 was increased. Following the administration of PP2, the increased p-Src and NMDAR-2B levels detected at 2 h after injury were decreased, and tissue injury and myelin basic protein expression were improved at 7 days after injury. The PP2 intervention improved the performance of injured rats on behavioral tests. In conclusion, we determined the patterns of p-Src expression after HI brain injury in immature rats and showed a relationship with the activated NMDA receptor. The inhibition of p-Src ameliorates neuropathological changes and damages neurological functions induced by HI injury.
Insights
Hypoxic-ischemic (HI) brain injury in neonatal rats involves Src family kinases (SFKs) and N-methyl-D-aspartate receptors (NMDARs). Inhibiting SFKs with PP2 reduced injury and improved neurological function in immature rats.
Area of Science:
- Neuroscience
- Biochemistry
- Pathology
Background:
- Neonatal hypoxic-ischemic (HI) brain injury is a significant clinical concern.
- Src family kinases (SFKs) are implicated in N-methyl-D-aspartate receptor (NMDAR) regulation and HI injury pathways.
Purpose of the Study:
- To investigate phospho-Src (p-Src) levels and NMDAR-SFK interactions in an immature rat HI brain injury model.
- To evaluate the therapeutic potential of SFK inhibition on HI-induced neuropathology and functional deficits.
Main Methods:
- Established a neonatal rat HI brain injury model (Rice-Vannucci).
- Assessed p-Src and NMDAR subunit NR2B phosphorylation via Western blotting and coimmunoprecipitation.
- Administered the SFK inhibitor PP2 and evaluated neuropathological changes (H&E, immunofluorescence) and neurological function (behavioral tests).
Main Results:
- p-Src levels exhibited dynamic changes post-HI, peaking at 2 hours with concurrent NR2B phosphorylation.
- PP2 treatment decreased elevated p-Src and NR2B phosphorylation.
- SFK inhibition improved histological outcomes, myelin basic protein expression, and behavioral performance in HI rats.
Conclusions:
- Dynamic changes in p-Src and NMDAR activation occur after neonatal HI brain injury.
- SFK inhibition, via PP2, mitigates neuropathological damage and functional impairments, suggesting SFKs as therapeutic targets.
