脂代谢相关三基因突变小鼠肝脏差异表达蛋白组研究

Differential Proteomic Analysis of Liver of Treble Lipid Metabolism Genes Mutant Mouse

应用双向电泳及质谱技术对5周龄三基因(apoELDLRLeprdbdb)联合突变小鼠和野生型小鼠肝组织的差异蛋白质进行比较研究,借此分析脂代谢相关三基因联合突变小鼠肝脏蛋白质表达特点,研究差异表达蛋白与血脂代谢紊乱和动脉粥样硬化的关系.在实验中检测到三基因联合突变小鼠和野生型小鼠肝脏中分别平均有(841±57)个和(1017±50)个蛋白点(n=3),两者的平均匹配率分别为71.9%,83.2%.三基因联合突变小鼠有140个蛋白点未能与野生型小鼠匹配,其中相差5倍以上的上调点和下调点分别为7个和39个.选取其中的6个点做质谱分析,鉴定为endoplasminprecursor(Grp_94)、酸性富亮氨酸核磷蛋白32家族成员A(acidicleucinrichnuclearphosphoprotein32familymemberA)、转铁蛋白前体、果糖二磷酸酶1、纤维连接蛋白前体、补体C3前体,纤维蛋白原Bβ多肽7种蛋白.该结果提示,差异表达的蛋白对三基因联合突变小鼠的血脂代谢紊乱和动脉粥样硬化发生发展过程起一定作用.

Two-dimensional gel electrophoresis and mass spectrometry have been used for analysis differential displayed proteomics of 5 week-old treble lipid metabolism genes mutant mice （apoE^-/-/LDLR^-/-/Lepr^db/db） and wild type mice. Comparative study of differential expression of protein profiles of livers between treble lipid metabolism genes mutant mice and wild type mice. The key protein related to atherosclerosis and dysfunction of lipid metabolism also characterized. Approximately （841±57） spots and （1 017±50） spots have been detected in treble lipid metabolism genes mutant mice livers （n = 3） and wild type mice livers （n = 3）. The average matched ratio was 71.9% and 83.2%. The differential expression analysis showed that there were matched spots between treble lipid metabolism genes mutant mice and wild type mice. Compared with wild identified by LTQ-ESI, e.g. endoplasmin precursor （Grp94）, acidic leucin-rich nuclear phosphoprotein 32 family member A, serotransferrin precursor, fructose bisphosphatase 1, fibronectin precursor, complement C3 precursor, fibrinogen B beta polypeptide. The protein profile of treble lipid metabolism genes mutant mice livers exhibited significant difference compared to that of wild type mice. The results implied that lipid metabolism relative polygenetic mutation contribute to the alteration of mouse liver protein expression profile, especially which lipid metabolism related and others participate in dysfunction in lipid metabolism during atherogenesis.