自发性肥胖型糖尿病大鼠血浆蛋白组学分析

Plasma Proteomic Analysis of Zucker Diabetic Fatty Rats

采用同位素标记相对和绝对定量(iTRAQ)技术对自发性肥胖型糖尿病(Zucker diabetic fatty,ZDF)大鼠的血浆蛋白组进行分析,比较其与Zucker瘦型(Zuker lean,ZL)大鼠血浆蛋白质表达的差异,从蛋白质水平分析ZDF大鼠作为2型糖尿病模式生物的特性。分别选取5周龄SPF(无特定病原体动物)级雄性ZDF大鼠和ZL大鼠饲养至12周,饲养过程中记录饮食饮水量,测量体重、血糖、胰岛素水平,处死后取血浆并去除血浆高丰度蛋白质,用iTRAQ试剂标记后进行质谱分析。ZDF大鼠在饲养过程中表现出2型糖尿病典型的多饮多食多尿特征,同时出现高血糖、高胰岛素血症和高胰岛素抵抗指数症状。ZDF大鼠与对照大鼠血浆中表达量差异超出2倍的蛋白质共有405个,其中C反应蛋白上调,一系列载脂蛋白、氧化应激相关蛋白质、凝血相关蛋白质均存在差异。ZDF大鼠还存在脂代谢异常,机体处于氧化应激状态,并且出现一定程度的心血管并发症。

In this paper,iTRAQ(isobaric tags for relative and absolute quantification)technology was used for proteomic analysis of Zucker diabetic fatty(ZDF)rat’s plasma.Compared with that of Zucker lean(ZL)rats,the differences in protein expression were characterized for type 2 diabetes at the protein level.Five-week-old SPF male ZDF rats were selected and were bred for 12 weeks.The dietary and water consumption were recorded,and body weight,blood glucose and insulin levels were measured.Plasma was taken after execution and the high abundance of plasma proteins were removed and labeled with iTRAQ reagents for mass spectrometry analysis.ZDF rats showed the typical characteristics of type 2 diabetes,such as excessive drinking,eating and urination,and symptoms of hyperglycemia,hyperinsulinemia and high insulin resistance index.There were 405 proteins with more than2-fold difference compared with controls in ZDF rat plasma,among which C-reactive protein was up-regulated,a series of apolipoproteins,oxidative stress-related proteins and coagulation-related proteins were differentially expressed.ZDF rats have abnormal lipid metabolism,a state of oxidative stress,and some degree of cardiovascular complications.