摘要
目的观察慢性弓形虫感染大鼠海马组织的蛋白质组的表达与变化。方法将6只雄性SD大鼠随机分成健康对照组和弓形虫感染组,每组3只。感染组每鼠腹腔注射4×107个弓形虫RH株速殖子,对照组注射等量生理盐水,于感染后第5天,尾静脉采血,吉氏染色验证大鼠感染情况。感染10周后,分离大鼠海马组织,抽提蛋白,并采用双向凝胶电泳分离,考马斯亮蓝染色后用PDQuest1.0软件进行图像分析,从胶中选取分离蛋白质点,应用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析,获取肽质量指纹图谱(PMF),采用Mascot软件检索SwissProt数据库鉴定蛋白质。结果血涂片结果证实,感染组大鼠均感染成功。感染组和对照组大鼠海马组织经双向凝胶电泳分离后,分别检出蛋白斑点数(311±19)个和(327±13)个,对2张电泳图进行匹配后发现有16个蛋白斑点在2组大鼠海马组织中的含量有明显变化,其中5个蛋白斑点在感染组大鼠海马蛋白双向电泳图谱中消失,11个蛋白斑点在2组大鼠海马组织中含量发生了3倍以上的变化,其中感染组上调4个,下调7个。有差异表达的16个蛋白斑点经质谱鉴定和数据库检索,发现9个蛋白,分别为磷酸激酶1、类烯醇化酶、谷氨酰合成酶、肌酸激酶、B型肌酸激酶、ATP合酶、线粒体顺乌头酸酶、肌动蛋白和未命名蛋白,前3个蛋白在感染组上调,其余6个下调。结论共获得9个慢性弓形虫感染大鼠与健康大鼠海马组织差异表达蛋白。
Objective To observe the proteome changes in the hippocampus tissue of rats with chronic Toxoplasma gondii infection. Methods Six male SD rats were randomly divided into control group and infection group. Each rat in infection group was intraperitoneally injected with 4×10^7 purified T. gondii tachyzoites. Rats in the control group received equivalent volumes of sterile normal saline. At the fifth day post-infection, blood samples were taken from the lateral tail vein and Giemsa staining of blood cells was performed to find Toxoplasma gondii. Rats were dissected at the 10th week post-infection, total protein in the hippocampus was separated by using two-dimensional gel electrophoresis （2-DE）. After Coomassie blue staining, the Image Analysis software was used to select and separate proteins on the gel. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry （MALDI-TOF-MS） was used for peptide mass fingerprint （PMF）. Proteins were identified by using Mascot software to search the MSDB and SwissProt databases. Results Microscopy examination of blood smears confirmed that the rats in infection group were all infected by T. gondii. The number of protein spots of rats from infection group and control group was 311±19 and 327±13, respectively. Compared with the control group, 5 protein spots disappeared, 4 protein spots were up-regulated and 7 were down- regulated in the infection group. The 9 differentially expressed protein spots were identified by MALDL-TOF-MS： phosphoglycerate kinase 1, similar to alpha-enolase, glutamine synthetase, creatine kinase, creatine kinase B-type, ATPsynthase, aeonitase 2, mitochondrial precursor, actin and an unnamed protein. The first three proteins were up-regulated and the other five proteins were down-regulated in infection group. Conclusion Nine differential expression proteins are found from the hippocampus tissue in rats chronically infected with T. gondii and normal SD rats.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2013年第6期454-457,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
江苏省卫生科研基金(No.X200739)~~
关键词
刚地弓形虫
大鼠
海马
蛋白质组
Toxoplasma gondii
Rat
Hippocampus
Proteomics
