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1.
Journal of Forensic Medicine ; (6): 107-111, 2011.
Article in Chinese | WPRIM | ID: wpr-983634

ABSTRACT

OBJECTIVE@#To explore the difference of expression of proteins between the serum and hippocampus after brain injury in rats.@*METHODS@#Male SD rats were used to establish brain injury model. The changes of proteins expression profile in serum and hippocampus at different time after brain injury were analyzed using weak cationic exchanger (WCX2) chips and immobilized metal affinity capture arrays-Cu (IMAC-Cu) chips by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry.@*RESULTS@#A total of 436 protein peaks were detected in serum and 346 protein peaks were detected in hippocampus using WCX2 chips. A total of 229 protein peaks were detected in serum and 345 protein peaks were detected in hippocampus using IMAC-Cu chips. The same 10 protein peaks were respectively detected in serum and hippocampus using WCX2 chips. The same 13 protein peaks were respectively detected in serum and hippocampus using IMAC-Cu chips.@*CONCLUSION@#The changes of protein expression profile in serum and hippocampus are obvious after closed brain injury and show a significant difference. The different proteins detected in serum and hippocampus using the same chip could be biochemical markers for determining brain injury.


Subject(s)
Animals , Male , Rats , Blood Proteins/analysis , Brain/pathology , Disease Models, Animal , Head Injuries, Closed/metabolism , Hippocampus/metabolism , Predictive Value of Tests , Protein Array Analysis/methods , Proteins/metabolism , Proteomics , Rats, Sprague-Dawley , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
2.
Chinese Journal of Pediatrics ; (12): 209-213, 2011.
Article in Chinese | WPRIM | ID: wpr-286129

ABSTRACT

<p><b>OBJECTIVE</b>To screen differentially expressed brain proteins with proteomic method in cerebral cortex of neonatal rats with congenital hypothyroidism.</p><p><b>METHOD</b>From the 13th day of gestation, pregnant Wistar rats from the experimental group were given intragastrically with 2.5 ml of 1% propylthiouracil daily. Cerebral cortex specimens were collected from the control and hypothyroidism neonatal rats. Two-directional electrophoresis (2-DE) was applied to analyze protein expression diversities between the euthyroid and hypothyroidism neonatal rat cerebral cortex. Protein spots with significantly different expression were screened and identified by mass spectrometry. Radioimmunoassay (RIA) was used to analyze serum FT(3), FT(4) levels of each groups.</p><p><b>RESULT</b>The body weight of hypothyroid neonatal rats were lower than those in the corresponding control group (t = -8.07, P < 0.01). The FT(3) levels of hypothyroid neonatal rats were lower than those in the corresponding control group (t = 5.39, P < 0.01). The FT(4) levels of hypothyroid neonatal rats were lower than those in the corresponding control group (t = 7.62, P < 0.01). Stable 2-DE maps of normal and CH neonatal rat were constantly obtained. The maps were analyzed by software. Seven protein spots with high reproducibility, high resolution and significantly different expression were chosen and identified by mass spectrometry, including collapsing response mediator protein 2, actin related protein 2/3 complex subunit 5, ubiquitin-conjugating enzyme E2-25K, ATP synthase subunit d, Cu-Zn superoxide dismutase, synuclein alpha, and nucleoside diphosphate kinase.</p><p><b>CONCLUSION</b>The value of this research is demonstrated here by the identification of several proteins known to be associated with nerve synapse structures formation, cell survival, metabolism, cell signal transduction, neural differentiation and nerve growth in the central nervous system. Furthermore this study identified several proteins except for collapsing response mediator protein 2 and Cu-Zn superoxide dismutase that have not previously been described in the literature and which may play an important role as either sensitive biomarkers of brain dysfunction caused by congenital hypothyroidism. In congenital hypothyroidism, brain development retardation may be related with some important processes, including abnormal synaptic formation, excess ROS production and apoptosis. The above-mentioned proteins may play critical roles in the processes, which provide valuable clues to clarify the pathogenesis of brain developmental disorders induced by congenital hypothyroidism.</p>


Subject(s)
Animals , Female , Pregnancy , Rats , Animals, Newborn , Metabolism , Brain , Metabolism , Cerebral Cortex , Metabolism , Congenital Hypothyroidism , Metabolism , Proteome , Proteomics
3.
Chinese Journal of Applied Physiology ; (6): 46-50, 2010.
Article in Chinese | WPRIM | ID: wpr-356220

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the morphology alterations and proteomics changes in primary astrocytes following fluid percussion injury.</p><p><b>METHODS</b>Primary cultures of astrocytes were prepared from cerebral hemispheres of 1-3 d-old SD rats, then, astrocytes were randomly divided into control group and injury group which were subjected to (0.2 +/- 0.01) MPa fluid percussion injury. The changes of protein expression pattern in astrocytes between injury and control groups were monitored with two dimensional gel electrophoresis.</p><p><b>RESULTS</b>Astrocytes' s abnonmalities of morphology after injury were apparent. The fluid percussion injury caused astrocytes edema, shrinkage, cell junction disconnection and necrosis at 2 h after injury. 24 h and 48 h after injury, most part of astrocytes's dendrites and soma became hypertrophy and showed a higher rate of cell proliferation. The dynamic proteomics changes were identified and total different 13 spots were detected in this study from the 2DE gels. The different displayed 5 spots were identified via MALDI-TOF: cofilin 1, destrin, phosphoglycerate mutase 1, NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 10, annexin 1.</p><p><b>CONCLUSION</b>The obvious alteration of morphology and protein expression pattern in primary cultured astrocytes could be induced after fluid percussion injury. The differential proteins detected were probably related to stress responses.</p>


Subject(s)
Animals , Female , Male , Rats , Animals, Newborn , Astrocytes , Cell Biology , Metabolism , Brain Injuries , Metabolism , Cerebral Cortex , Cell Biology , Primary Cell Culture , Methods , Proteins , Metabolism , Proteome , Proteomics , Methods , Rats, Sprague-Dawley , Stress, Physiological , Physiology
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