慢性间歇性缺氧大鼠肾脏组织蛋白质组表达变化的初步研究  

Preliminary study on changes of expressive proteome in renal tissue of chronic intermittent hypoxia rats

作　　者：张春燕[1] 杨烨[1] 陈燕[1,2] 李洪[1] 赵春玲[2] 

机构地区：[1]泸州医学院生物化学教研室,四川省泸州市646000 [2]泸州医学院生理教研室,四川省泸州市646000

出　　处：《中华肾脏病杂志》2011年第8期602-605,共4页Chinese Journal of Nephrology

基　　金：四川省教育厅基金（2007ZC027）

摘　　要：目的比较慢性间歇性缺氧（CIH）大鼠与正常大鼠肾脏组织蛋白质双向电泳图谱，寻找和鉴定CIH大鼠肾脏组织中的差异蛋白表达谱。方法以固相pH梯度等电聚焦为第一向和垂直十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）为第二向，分别对正常对照大鼠（n=5）和CIH大鼠（n=5）肾脏组织蛋白质样品进行二维分离。二维电泳（2-DE）图谱经ImageMaster 2D Platinum 5．0软件分析，选取CIH组4个差异蛋白点，用基质辅助激光解吸附离子化飞行时间质谱（MALDI—TOF—MS）进行鉴定。结果通过对2-DE图谱蛋白斑点的匹配及对比分析，与CIH相关的差异表达蛋白斑点为112个。3个蛋白斑点经质谱鉴定为有意义的差异表达，下调的差异点1个，即线粒体ATP合成酶8亚基；上调的差异点2个，分别为儿茶酚-O-甲基转移酶、脱嘌呤脱嘧啶核酸内切酶。结论初步鉴定了与大鼠肾脏组织CIH损伤相关的差异蛋白，其表达的变化可能通过多种途经影响肾脏功能，这为CIH导致肾功能损害的发病机制研究提供了新的思路。Objective To obtain two-dimensional gel electrophoresis maps of the renal tissue proteins of normal and chronic intermittent hypoxia （CIH） rats for identifying the diferentially expressed proteins in the CIH rats. Methods CIH rat models were established, and the proteins in the renal tissue underwent two-dimensional gel electrophoresis with immobiline pH gradient isoeleclric focusing as the first dimension and vertical SDS-PAGE as the second dimension. Analysis of 2-DE maps was used to determine differential expression of proteins between the two groups by ImageMaster 2D Platinum 5.0, and four protein spots expressed differently were picked up for further identification by MALDI-TOF-MS. Results Matched and compared with those of control group, 112 protein spots were determined with differently expressive levels in CIH group. By MALDI-TOF-MS, three protein spots of them were identified as ATP synthase delta subunit of mitochondrial precursor, catechol O-methyltransferase, apurinic/apyrimidinic endonudease. Conclusions There is obvious difference in expressive proteomes in renal tissue between normal and chronic intermittent hypoxia rats. The functions of those identified proteins are involved in cellular energy metabolism, apoptosis, signal transduction, anti-cell injury and hormone metabolism, so that proteomics can serve as a new approach in the study of obstructive sleep apnea-hypopnea syndrome to discover new therapeutic targets.

关 键 词：蛋白质组学 睡眠呼吸暂停 阻塞性 蛋白质组 慢性间歇性缺氧 肾脏 双向凝胶电泳 

分 类 号：R766[医药卫生—耳鼻咽喉科]