突触后密度蛋白-93在大鼠脊髓损伤后的表达变化  

EXPRESSION OF POSTSYNAPTIC DENSITY PROTEIN-93 IN INJURIED SPINAL CORD OF RAT

作　　者：高尚锋[1] 程纯[2] 陈梦玲[2] 赵剑[3] 李欣[2] 牛淑琼[1] 严斌[3] 沈爱国[1] 

机构地区：[1]南通大学江苏省神经再生重点实验室,南通226001 [2]南通大学基础医学院微生物与免疫学教研室,南通226001 [3]南通大学附属医院骨科,南通226001

出　　处：《解剖学报》2007年第4期405-409,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30300099);江苏省自然科学基金资助项目(BK2003035)(BK2006547);江苏省高校自然科学研究项目(03KJB180109;04KJB320114);江苏省社会发展科技指导性计划项目(BS2004526);江苏省"六大人才高峰"第二批资助项目资助

摘　　要：目的探讨突触后密度蛋白-93(PSD-93)在脊髓损伤(SCI)后的表达变化以及定位情况。方法使用改良Allen法制备大鼠急性脊髓损伤模型,致大鼠脊髓(T9、T10)重度撞击伤。采用实时定量PCR(Real-time PCR)和Western blotting测定损伤后各时间段PSD-93 mRNA及其蛋白水平在脊髓中的表达变化。采用免疫荧光双标方法检测PSD-93在正常脊髓中的分布以及损伤后的定位改变。结果PSD-93 mRNA水平在SCI后呈现逐渐下降的趋势,在3d降到最低水平;蛋白水平则在SCI后显著上调,于1d达高峰。免疫荧光双标显示,PSD-93在SCI后除与神经型一氧化氮合酶(nNOS)存在着共定位之外,还高表达于损伤局部活化的小胶质细胞和星形胶质细胞。结论脊髓损伤后PSD-93在基因和蛋白水平呈现明显的时相变化,并且与nNOS、活化的胶质细胞存在共定位,提示PSD-93参与了脊髓继发性损伤过程。Objective To study the expression and distribution of postsynaptic density pretein-93 （PSD-93） after spinal cord injury （SCI）. Methods With improved Allen＇ s method, acute rats SCI models were established. Real-time PCR and Western blotting were used to detect the changes of PSD-93 mRNA and protein expression after SCI. The changes of PSD-93 localization after injury were investigated by immunofluorescence double staining. Results The expression of PSD-93 mRNA had a gradually decrease trend after SCI. The minimal mRNA was present at 3days; whereas the protein levels up-regulated significantly and peaked at lday after injury. Immunofluorescence double staining indictated that PSD-93 was highly expressed on the activated microglias and astrocytes except that it co-localized with nNOS in the impaired spinal cord. Conclusion The time course of PSD-93 and its mRNA is vary significantly after SCI, and colocalized with nNOS and activated glias. These results indicate that PSD-93 may participate in the following response after SCI.

关 键 词：突触后密度蛋白-93 脊髓损伤 神经型一氧化氮合酶 实时定量PCR 免疫印迹法 免疫荧光 大鼠 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]