自发性高血压大鼠丙泊酚麻醉后海马组织的蛋白质组学研究

Study on proteomic profiling of hippocampus tissues of spontaneously hypertensive rats after propofol anesthesia

目的观察丙泊酚麻醉对自发性高血压大鼠海马组织蛋白质表达的影响及其与认知功能的关系。方法 40只自发性高血压大鼠随机分为实验组(n=20)和对照组(n=20)。实验组经腹腔注射丙泊酚100 mg/kg,1 h和2 h后追加首剂量的1/2,维持麻醉3 h;对照组采用同样方法和剂量给予生理盐水作为对照。分别于给药后3、24、72 h及7 d时,随机选取两组大鼠各5只,取海马组织提取总蛋白样品。采用双向凝胶电泳分离蛋白样品,分析差异表达蛋白点(差异>2倍的蛋白质点),基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)鉴定。通过数据库检索分析差异蛋白的主要功能和参与的生物学过程。结果获得差异表达的蛋白质点共有84个,其中68个经MALDI-TOF-MS成功鉴定,去冗余后经蛋白质数据库搜寻确定了47个差异表达蛋白。以蛋白的低表达为主,持续到麻醉后第7日。多种生物学过程(涉及能量代谢、线粒体功能、蛋白折叠和囊泡转运等)受到影响。结论自发性高血压大鼠丙泊酚麻醉后海马组织蛋白发生多维、动态改变,部分蛋白的差异表达可能参与认知功能损害的分子机制,具体作用有待进一步深入探讨。

Objective To investigate the effects of propofol anesthesia on the expressions of hippocampus protein and it＇s relationship with the cognitive function of spontaneously hypertensive rats. Methods Forty spontaneously hypertensive rats were randomly divided into the experiment group （ n =20） and control group （n =20）. The rats of experiment group were given 100 mg/kg of propofol by peritoneal injection for anesthetic induction and the anesthesia was maintained for 3 h by administering half dose of inductive propofol after 1 h and 2 h. The control group was treated by saline with the same procedure and volume as to propofol injection. Five rats in each group were randomly chosen and decapitated at 3 h, 24 h, 72 h, and on 7 d after anesthesia. Hippocampus tissues were immediately removed to an ice-board and subjected to global protein expression profiling based on two-dimensional gel electrophoresis. Spots expressed with more than two-fold changes were cut out for analysis by the MALDI-TOF-MS. The major functions and involved biological processes of differential proteins were analyzed by database retrievals. Results Eighty-four differentially expressed proteins were detected, among which sixty eight proteins were successfully identified by the MALDI-TOF MS and 47 proteins were determined by retrievals of protein databases after duplicating ones were removed. Most proteins were low expressed and maintained till the 7 d after anesthesia. Many biological processes （involving the energy metabolism, mitochondrial function, protein folding, and vesicle trafficking, etc. ） were affected. Conclusion The influence of propofol anesthesia results in multi-dimensional and dynamic changes of hippocampnsin ofspontaneously hypertensive rats. Differential expressions of some proteins may involve the molecular mechanism of impairing the cognitive function, but its actual effects need further investigation.