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目的检测脑挫伤修复过程中突触后致密蛋白(postsynaptic density protein 95,PSD-95)的表达,探讨其变化规律与损伤时间的相关性。方法雄性SD大鼠50只,随机分为8个实验组,1个对照组,每组5只。制作大鼠脑挫伤模型,于伤后3、6、12h和3、5、7、10d取脑组织,应用免疫组织化学技术和免疫蛋白印迹(Western blot)方法检测脑挫伤后不同时间脑组织中PSD-95的表达变化。结果对照组脑组织仅有少量PSD-95阳性细胞;实验组中,伤后3h、6h组脑组织出现较多PSD-95阳性细胞,12h组阳性细胞数量持续升高,伤后1d阳性细胞数下降,5d后又升高并达到高峰,7d、10d恢复;计算阳性率,统计分析结果显示,阳性细胞数与相邻上组比较,存在显著性差异。Western blot结果:挫伤后,3～12h表达量上升,1d下降,随后又逐步上升,5d达到高峰,7d、10d下降;应用Fluorchem V2.0 Stand Alone软件获取感光条带的平均灰度值,经统计分析,各组与相邻上组比较,存在显著性差异。结论大鼠脑损伤后损伤周边区PSD-95呈现升高→下降→再升高→再下降的表达规律,对损伤时间推断有一定的参考意义。
Objective To detect the expression of postsynaptic density protein 95 (PSD-95) in the process of brain contusion repair and to explore the relationship between the changes and the injury time. Methods Fifty male SD rats were randomly divided into 8 experimental groups and 1 control group, with 5 rats in each group. Brain contusion models were made in rats. Brain tissue was harvested at 3, 6, 12h and 3, 5, 7 and 10 days after injury. Immunohistochemistry and Western blotting were used to detect brain injury at different time points PSD-95 expression changes. Results There were only a few PSD-95 positive cells in the control group. In the experimental group, more PSD-95 positive cells appeared in the brain tissue at 3h and 6h after injury, while the number of positive cells in the 12h group continued to increase. And then increased and peaked on the 5th day, and resumed on the 7th day and the 10th day. The positive rate was calculated. The statistical analysis showed that the number of positive cells was significantly different from the adjacent upper group. Western blot results: After contusion, the expression level increased from 3 to 12 h, decreased on the 1st day, then gradually increased, reached its peak on the 5th day, decreased on the 7th day and the 10th day. The average gray value of the photosensitive band was obtained by Fluorchem V2.0 Stand Alone software. Statistical analysis, each group compared with the adjacent group, there are significant differences. Conclusion The expression of PSD-95 in the peripheral area of injured area after traumatic brain injury in rats is increased, decreased, then increased and then decreased. It is of some reference significance to infer the damage time.