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摘要
BACKGROUND: Studies have reported the combined use of two-dimensional gel electrophoresis and mass spectrometry to detect differentially expressed proteins in the rat brainstem following brain injury. However, the detected differential proteins often exhibit low sensitivity and high relative molecular weight. Although protein chip technology is thought to compensate for these inadequacies, no related studies or results have been reported.
OBJECTIVE: To propose the application of weak cation exchange protein chips in combination with mass spectrometry for determining protein expression profiles and characteristics in the brainstem following closed brain injury.
DESIGN, TIME AND SETTING: Randomized, controlled, animal experiments utilizing proteomics were performed from June 2007 to December 2008 in the Proteomics Laboratory, Medical College of Chinese People's Armed Police Force.
MATERIALS: Weak cation exchange 2 protein chip, Ciphergen Proteinchip System (PBS-IIC).
METHODS: A total of 72 rats were randomly assigned to two groups: sham-surgery (n = 12) and injury (n = 60). A closed traumatic brain injury model caused by falling object was replicated in the injury group, which was then subdivided into five subgroups according to different time points after injury: 4, 8, 12, 24, and 48 hours, with 12 rats in each subgroup. In the sham-surgery group, only the skin was removed and the stainless steel pad was fixed to the skull.
MAIN OUTCOME MEASURES: The brain injury rats were sacrificed at 4, 8, 12, 24, and 48 hours after injury, respectively, and the control rats were sacrificed at 24 hours. Pathological changes in the brainstem were determined using hematoxylin-eosin staining, and differential protein expression in the brainstem was detected using a weak cation exchange 2 protein chip and protein chip reader.
RESULTS: In the sham-surgery group, cells appeared normal. However, in the brain injury group, some brainstem neurons exhibited pyknosis, with reduced numbers of Nissl bodies in the cytoplasm, swollen cell bodies and nuclei, irregular staining in the cytoplasm, and decreased numbers of neurons. Results from weak cation exchange 2 protein chip detection demonstrated that, compared with the sham-surgery group, the expression profiles of 2 proteins were altered in the brainstem of the injury group. At 12, 24, and 48 hours after injury, expression was increased (P < 0.01). The mass charge ratio (M/Z) of 7 862 differentially expressed proteins was greater in the sham-surgery group compared with 12 and 24 hours after injury (P < 0.05).
CONCLUSION: The combined method of weak cation exchange 2 protein chip and mass spectrometry detected differential protein expression in the brainstem following closed brain injury in the rats, which suggested that closed brain injury induced altered protein expression profiles in the brainstem.
- 出版日期2010-3-15
- 单位中国人民武装警察部队后勤学院
