丙泊酚对糖尿病大鼠海马组织蛋白质表达影响的蛋白组学研究

Proteomics study of effects of propofol on expression of proteins in hippocampus tissues of diabetic rats

目的采用蛋白组学方法研究丙泊酚对2型糖尿病Goto-Kakizaki(GK)大鼠海马组织蛋白质表达的影响。方法将清洁级GK大鼠随机分为对照组(n=12)、丙泊酚麻醉组(n=12)和参数组(n=5),丙泊酚组和参数组腹腔注射丙泊酚100 mg/kg,在1 h和2 h后追加剂量50 mg/kg,维持麻醉3 h,对照组采用同样方法给予等体积生理盐水。丙泊酚组分别在麻醉结束后3、24、72 h和1周4个时间点随机选取3只大鼠,取海马组织,双向电泳法分离蛋白后进行凝胶扫描和图像分析,与相应时间点的对照组进行比较,选取4倍差异点进行质谱分析。观察参数组丙泊酚给药后0.5、1.5和2.5 h时间点血气分析指标的变化。结果与对照组相比,丙泊酚组麻醉后3 h发现7个蛋白点下调,24 h有3个蛋白点下调,72 h有2个蛋白点下调,1周则出现2个蛋白点上调,经质谱鉴定发现14个差异表达蛋白,涉及氧化还原、能量代谢、突触信息传递、细胞骨架和运动、细胞凋亡等生物学过程。血气分析指标监测结果显示:参数组丙泊酚给药后0.5 h与2.5 h时点的pH值和氧分压(PaO2)比较差异有统计学意义(P<0.05),但均在正常生理指标范围内。结论丙泊酚麻醉后不同时间点GK大鼠海马组织多种蛋白质的表达出现异常,其中海马胆碱能神经刺激肽前体蛋白、电压依赖性阴离子选择性通道蛋白1等蛋白质与认知功能密切相关。

Objective To investigate the effects of propofol on the expression of proteins in hippocampus tissues of Goto-Kakizaki(GK) rats with type 2 diabetes mellitus by proteomics method.Methods GK rats were randomly allocated to propofol group(n=12),control group(n=12) and parameter group(n=5).Rats in propofol group and parameter group were anesthetized with propofol intraperitoneally under an initial bolus injection(100 mg/kg),followed by supplemental doses of 50 mg/kg 1 h and 2 h later,and the anesthesia maintained for 3 h.Rats in control group were injected with the same amount of normal saline.Rats in propofol group were sacrificed 3 h,24 h,72 h and 1 week after anesthesia respectively,and hippocampus tissues were obtained and subjected to global protein expression profiling based on SDS-PAGE.Expression factors were compared with results from control group at each time point,and spots expressed more than four-fold change were cut out for mass spectrometry.Rats in parameter group were selected for blood gas analysis 0.5 h,1.5 h and 2.5 h after administration of propofol.Results Compared with control group,down-regulated spots were found 3 h(7 spots),24 h(3 spots) and 72 h(2 spots) after anesthesia,2 up-regulated spots were detected at the time point of 1 week,and 14 differentially expressed proteins were identified in relation to redox reactions,energy metabolism,synaptic transmission,cytoskeleton and movement,and cell apoptosis in propofol group.Blood gas analysis indicated that there were significant differences in pH and PaO2 between the time points of 0.5 h and 2.5 h after administration of propofol in parameter group(P<0.05),while both were in the normal physiological range.Conclusion Propofol anesthesia may result in changes of expression of proteins in hippocampus tissues of GK rats,and proteins such as hippocampal cholinergic neurostimulating peptide precursor and voltage-dependent anion-selective channel protein 1 are associated with cognitive function.