摘要
目的观察复合营养素和植物提取物组方干预对老龄大鼠额叶皮层蛋白质组表达的影响。方法选取15月龄Wistar大鼠,雄性14只,雌性16只,随机分为正常对照组和营养干预组,每组雄性7只,雌性8只。营养干预组大鼠饲喂添加复合营养素的自制饲料,同时灌胃植物提取物混合液,而对照组饲喂基础饲料,同时灌胃蒸馏水。灌胃剂量为4ml/(kgbw·d)。实验期10w。建立大鼠额叶皮层蛋白质组研究方法,二维凝胶电泳(2-DE)检测大鼠额叶皮层蛋白质表达的变化,利用辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)和Mascot数据库检索鉴定表达差异的蛋白,Westernblot检测额叶皮层stathmin1蛋白的表达。结果与正常对照组比较,营养干预组老龄大鼠额叶皮层2-DE图谱中11个蛋白点含量发生变化,其中丙酮酸激酶、丙酮酸脱氢酶E1α1、天门冬氨酸转氨酶、线粒体肌酸激酶1、电压依赖性阴离子通道2、磷酸丙糖异构酶1等6种蛋白质含量降低,而过氧化物氧化还原酶2、线粒体H-ATP合酶d亚单位、stathmin1、Cu,Zn-SOD等4种蛋白质含量升高。Westernblot验证了额叶皮层stathmin1蛋白的表达变化。结论复合营养素和植物提取物组方改善老龄大鼠认知功能的作用机制可能与额叶皮层中认知相关蛋白质表达模式的改变有关。
Objective To investigate the effects of nutritional intervention on the differentially expressed protein in frontal cortex in aged rats by proteomics study. Method Fourteen male and sixteen female Wistar aged rats were randomly divided into intervened group and control group, seven male and eight female in each. The rats in intervened group were fed with diets supplemented with combined nutrients and mixed plant extract administered ig at 4 ml/（kg bw·d）, while the rats in control group were fed with examined basic diets and water. The experiment lasted 10 w. Protein expression in frontal cortex of aged rats was examined by two dimensional gel electrophoresis（2-DE）. The differentially expressed proteins were identified by MALDI-TOF-MS and Mascot software. Western blot was used to confirm the differentilly expressed protein stathmin 1. Results Eleven protein spots with changes of contents were observed in two dimensional gel electrophoresis maps and ten of them were identified successfully. Among the identified proteins, six proteins decreased in frontal cortex of dietary supplemented rats including pyruvate kinase, pyruvate dehydrogenase E1 alpha 1, aspartate transaminase, mitochondrial creatine kinase 1, voltage-dependent anion channel 2 protein and triosephosphate isomerase 1, while the other four proteins increased, including peroxiredoxin 2, subunit d of mitochondrial H-ATP synthase, stathmin 1 and Cu-Zn superoxide dismutase. The results of Western blot was consistent with that of proteomic observation.Conclusion The mechanism of the combined nutrients and plant extractives might correlate with the changes of expression pattern of several cognition related proteins in frontal cortex.
出处
《营养学报》
CAS
CSCD
北大核心
2009年第6期521-526,共6页
Acta Nutrimenta Sinica
基金
中国营养学会科研基金(No2004-07)
天津市应用基础及前沿技术研究计划(No09JCYBJC12900)
关键词
蛋白质组
衰老
认知
营养干预
大鼠
proteomics aging cognition nutritional intervention
